EP1404652A1 - A novel alkaloid derivative and a pharmaceutical composition containing the same - Google Patents

A novel alkaloid derivative and a pharmaceutical composition containing the same

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Publication number
EP1404652A1
EP1404652A1 EP02733537A EP02733537A EP1404652A1 EP 1404652 A1 EP1404652 A1 EP 1404652A1 EP 02733537 A EP02733537 A EP 02733537A EP 02733537 A EP02733537 A EP 02733537A EP 1404652 A1 EP1404652 A1 EP 1404652A1
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EP
European Patent Office
Prior art keywords
compound
oxo
benzo
heptarene
tetrahydro
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EP02733537A
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German (de)
French (fr)
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EP1404652B1 (en
EP1404652A4 (en
Inventor
Wan Joo Kim
Kyoung Soo Kim
Myung Hwa Kim
Jong Yek Park
Jung Min Jang
Jae Won Choi
Dong Hoo Kim
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Chemtech Research Inc
KT&G Corp
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Chemtech Research Inc
KT&G Corp
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C69/00—Esters of carboxylic acids; Esters of carbonic or haloformic acids
    • C07C69/76—Esters of carboxylic acids having a carboxyl group bound to a carbon atom of a six-membered aromatic ring
    • C07C69/78—Benzoic acid esters
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
    • A61P21/02—Muscle relaxants, e.g. for tetanus or cramps
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C203/00—Esters of nitric or nitrous acid
    • C07C203/02—Esters of nitric acid
    • C07C203/10—Esters of nitric acid having nitrate groups bound to carbon atoms of six-membered aromatic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C233/00—Carboxylic acid amides
    • C07C233/01—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C233/30—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by doubly-bound oxygen atoms
    • C07C233/32—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by doubly-bound oxygen atoms with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by a carbon atom of a ring other than a six-membered aromatic ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C233/00—Carboxylic acid amides
    • C07C233/01—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C233/34—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by amino groups
    • C07C233/41—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by amino groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by a carbon atom of a ring other than a six-membered aromatic ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C233/00—Carboxylic acid amides
    • C07C233/64—Carboxylic acid amides having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings
    • C07C233/76—Carboxylic acid amides having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by doubly-bound oxygen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C235/00—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms
    • C07C235/02—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton
    • C07C235/04—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C235/14—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton the carbon skeleton being acyclic and saturated having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a ring other than a six-membered aromatic ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C235/00—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms
    • C07C235/42—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings and singly-bound oxygen atoms bound to the same carbon skeleton
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C271/00—Derivatives of carbamic acids, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups
    • C07C271/06—Esters of carbamic acids
    • C07C271/08—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms
    • C07C271/24—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atom of at least one of the carbamate groups bound to a carbon atom of a ring other than a six-membered aromatic ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C323/00—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups
    • C07C323/23—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and nitrogen atoms, not being part of nitro or nitroso groups, bound to the same carbon skeleton
    • C07C323/39—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and nitrogen atoms, not being part of nitro or nitroso groups, bound to the same carbon skeleton at least one of the nitrogen atoms being part of any of the groups, X being a hetero atom, Y being any atom
    • C07C323/40—Y being a hydrogen or a carbon atom
    • C07C323/41—Y being a hydrogen or an acyclic carbon atom
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C323/00—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups
    • C07C323/23—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and nitrogen atoms, not being part of nitro or nitroso groups, bound to the same carbon skeleton
    • C07C323/39—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and nitrogen atoms, not being part of nitro or nitroso groups, bound to the same carbon skeleton at least one of the nitrogen atoms being part of any of the groups, X being a hetero atom, Y being any atom
    • C07C323/40—Y being a hydrogen or a carbon atom
    • C07C323/42—Y being a carbon atom of a six-membered aromatic ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C49/00—Ketones; Ketenes; Dimeric ketenes; Ketonic chelates
    • C07C49/587—Unsaturated compounds containing a keto groups being part of a ring
    • C07C49/753—Unsaturated compounds containing a keto groups being part of a ring containing ether groups, groups, groups, or groups
    • C07C49/755—Unsaturated compounds containing a keto groups being part of a ring containing ether groups, groups, groups, or groups a keto group being part of a condensed ring system with two or three rings, at least one ring being a six-membered aromatic ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
    • C07D307/77—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D307/93—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom ortho- or peri-condensed with carbocyclic rings or ring systems condensed with a ring other than six-membered
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2603/00—Systems containing at least three condensed rings
    • C07C2603/02—Ortho- or ortho- and peri-condensed systems
    • C07C2603/04—Ortho- or ortho- and peri-condensed systems containing three rings
    • C07C2603/30—Ortho- or ortho- and peri-condensed systems containing three rings containing seven-membered rings
    • C07C2603/34—Benzoheptalenes; Hydrogenated benzoheptalenes

Definitions

  • the present invention relates to a novel alkaloid derivative and a pharmaceutical composition containing the same. More particularly, the present invention relates to a novel colchicine derivative having anticancer, anti-proliferous and anti-inflammatory effects and immunosuppressive and muscle relaxing functions, and pharmaceutically acceptable salts thereof, a pharmaceutical composition containing the same as an effective component, and methods for preparing the same.
  • Colchicine is a pseudo-alkaloid widely used for treatment of gout and is used only for short-term therapeutic treatment due to its toxicity. However, colchicine has been reported to exhibit a very fast and specific therapeutic effect on gout, as described in the Alkaloids, 1991, vol. 41, 125-176, U.S. patent No. 4,533,675, and so on.
  • colchicine inhibits formation of mitotic spindle, thereby suppresses cell division, leading to activation of anticancer and anti-proliferous effects. Also, continuous research into colchicine applications has been carried out and a variety of colchicine derivatives have been synthesized up to now, as described in U.S. patent No. 3,222,253, U.S. patent application serial No. 00/608073A, WO
  • 91/02084, and so on are examples of demecolcine.
  • demecolcine has been used for treatment of leukemia.
  • colchicine can be used for treatment of psoriasis or rheumatoid arthritis and has an amyloidosis inhibitory effect and an anti-inflammatory effect (Arch. Dermatol. 1982, Vol. 118, July, pp 453-457).
  • thiocolchicoside which is one of colchicine derivatives, is widely used for treatment of skeletal muscle contracture and inflammation.
  • a first feature of the present invention is to provide a novel colchicine derivative having anticancer, anti-proliferous and anti-inflammatory effects and immunosuppressive and muscle-relaxing functions, and pharmaceutically acceptable salts thereof.
  • a second feature of the present invention is to provide methods for preparing the colchicine derivatives.
  • a third feature of the present invention is to provide a pharmaceutical composition containing the colchicine derivative and pharmaceutically acceptable salts thereof as an effective component.
  • N(R ) 2 , R 3 and R- t are independently hydrogen or a methyl group
  • R 5 is hydrogen, a methyl group or CH 2 X 4 R 7 , wherein R ⁇ and R 7 are independently hydrogen or a lower alkyl, and X, Xi, X 2 , X and X 4 are independently O or S;
  • R 2 is N(Re)C(X ⁇ )-A, X 2 C(X -A, N(Re)-A, N(A) 2 - or X 2 -A, R, is N(R 6 )COCH 3 , N(Re)COCF 3 , or NHC(O)OR 8
  • R 3 and R 4 are independently hydrogen or a methyl group
  • R 5 is hydrogen, a methyl group or CH 2 X 4 R , wherein R ⁇ and R 7 are independently hydrogen or a lower alkyl, R 8 is a lower alkyl, alkenyl, or substituted or unsubstituted aryl, and
  • R ⁇ and R 7 are independently hydrogen or a lower alkyl
  • R 8 is a lower alkyl, alkenyl, or substituted or unsubstituted aryl
  • R 5 is CH 2 X 2 C(X,)-A
  • R is N(R 6 )COCH 3 , N(R 6 )COCF 3 or NHC(O)OR 8
  • R 2 is X 3 R 7 or N(R 7 ) 2
  • R 3 and ⁇ are independently hydrogen or a methyl group
  • R$ and R 7 are independently hydrogen or a lower alkyl
  • R 8 is a lower alkyl, alkenyl, or substituted or unsubstituted aryl
  • X, X ⁇ ,X 2 and X are independently O or S
  • A is represented by the formula (a), (b), (c), (d), (e), (f), (g), (
  • Yj is a Ci to C ⁇ 0 straight chain or branched alkyl, preferably, a C 2 to C 5 straight chain or branched alkyl or a substituted C 5 to C 7 cycloalkyl;
  • Hal is halogen, for example, F, Cl, Br or I
  • R 9 is hydrogen or a lower alkyl;
  • ni is an integer from 1 to 6, preferably from 2 to 4;
  • n 2 and n 3 are independently an integer from 1 to 5, preferably from 1 to 3; is an integer from 0 to 3; and
  • n 5 is an integer from 1 to 6.
  • a pharmaceutical composition containing the colchicine derivatives and pharmaceutically acceptable salts thereof as effective components, the pharmaceutical composition having anticancer, anti-proliferous and anti-inflammatory effects and immunosuppressive and muscle-relaxing functions.
  • methods for preparing the colchicine derivative represented by the formula (I) according to the reaction schemes 1, 2, 3, 4 and 5.
  • the colchicine derivatives of the formula (I) are represented as the formula (la), (lb), (Ic), (Id), or (Ie):
  • R ⁇ is hydrogen or a lower alkyl
  • X ⁇ and X 2 are independently O or S
  • Hal, Hah, and Hal 2 may be the same or different halogens
  • L is a leaving group selected from methanesulfonyl, ?-toluenesulfonyl or triflate
  • P represents a general hydroxy-protecting group including methoxymethyl or t-butyldimethylsilyl
  • R 10 is hydrogen or a lower alcohol of to C 3
  • Y is represented by the general formula (a'Mb'Mc'Md ⁇ or ⁇ ):
  • Yi represents a Ci to C ⁇ 0 straight chain or branched alkyl, preferably, a C 2 to C 5 straight chain or branched alkyl, or a substituted C 5 to C 7 cycloalkyl
  • R 8 represents hydrogen or a lower alkyl
  • ni is an integer from 1 to 6, preferably an integer from 2 to 4
  • n 2 and n 3 are independently an integer from 1 to 5, preferably an integer from 1 to 3
  • r is an integer 0 to 3
  • n 5 is an integer from 1 to 6.
  • FIG. 1 is a graph showing the immonosuppressive effect of a colchicine derivative tested by mixed lymphocyte reaction (MLR);
  • FIG. 2 is a graph showing the immonosuppressive effect of cyclosphorine A as a positive control sample in an immunosuppressive effect test using a BALB/c mouse spleen;
  • FIG. 3 is a graph showing the immonosuppressive effect of a colchicine derivative according to the present invention in an immunosuppressive effect test using a BALB/c mouse spleen.
  • a lower alkyl represents a saturated Ci to C 6 , preferably, Cx to C 4 straight chain or branched hydrocarbon.
  • colchicine derivative of the formula (1) and its pharmaceutically acceptable salt according to the present invention include:
  • (+)-3-nitrooxymethyl-benzoic acid 1, 2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl ester;
  • N-(10-dimethylamino-l,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-7 -yl)-4-nitrooxy-butyrilamide examples include, but are not limited to, salts with inorganic bases such as sodium, potassium, magnesium or calcium and salts with organic bases such as ammonium, lysine, ethanolamine, N,N'-dibenzylethylenediamine and angelic acid Preparation methods of the invention will now be described in more detail.
  • Method 1 for preparing a compound of the formula (la) first, a compound of the formula (H) is reacted with a compound of the formula (III) to produce a compound (IN).
  • the reaction can be carried out without using a base, but is generally carried out in the presence of a base used for amidation.
  • Preferred examples of the base include pyridine, triethylamine, diethylisopropylamine and dimethylphenylamine, and a phase transfer catalyst such as sodium hydrocarbonate or benzyltriethylammonium chloride.
  • the reaction can be carried out without using a solvent, but is advantageously carried out in the presence of a solvent that does not adversely affect the reaction.
  • reaction temperature is not specifically limited and is generally carried out at reduced or elevated temperature, preferably at room temperature.
  • the compound of the formula (IV) is subjected to nitration to convert it into a compound of the formula (la).
  • Compounds capable of nitrating halogen are generally used for the reaction, and examples of such compounds include AgNO 3 and t-butylammonium nitrate (B11 4 NNO 3 ).
  • the reaction is preferably carried out in the presence of a solvent that does not adversely affect the reaction, and examples of such solvent include chloroform, acetonitrile, a mixed solution of acetonitrile and water, dichloromethane, and the like.
  • the reaction temperature is not specifically limited and is generally carried out at reduced or elevated temperature, preferably at room temperature.
  • the compound of the formula (la) can be synthesized by reacting the compound of the formula (U) with a compound of the formula (V) to produce a compound (VI), followed by converting it into the compound (la).
  • the conditions for reacting the compound of the formula (H) with the compound of the formula (V) are the same as those for amidation like in the reaction between the compound of the formula (H) and the compound of the formula (III).
  • the reaction is generally carried out under nitrating conditions of alcohol.
  • nitric acid and sulfuric acid, dinitrogen pentaoxide (N 2 O 5 ) and aluminum chloride HI, potassium nitrate and boron trifluoride (BF 3 ), acetylnitrate, etc. may be used, most preferably nitric acid and acetic anhydride (Ac 2 O) are used.
  • the reaction is preferably carried out in the presence of a solvent that does not adversely affect the reaction, and examples of the solvent used include chloroform, dichloromethane, and the like.
  • the reaction temperature is not specifically limited and is generally carried out at reduced or elevated temperature, preferably at room temperature.
  • the compound of the formula (la) can also be synthesized by performing nitration a compound of the formula (VH) prepared by converting hydrogen of alcohol in the compound of the formula (VI) into a leaving group such as methane sulfonyl, 7-toluene sulfonyl or triflate.
  • a compound of the formula (VH) prepared by converting hydrogen of alcohol in the compound of the formula (VI) into a leaving group such as methane sulfonyl, 7-toluene sulfonyl or triflate.
  • the reaction is generally carried out under nitrating conditions.
  • t-butylammonium nitrate (Bu 4 NNO 3 ) and nitric acid, nitric acid and silver nitrate, or potassium nitrate is used.
  • the reaction is preferably carried out in the presence of a solvent that does not adversely affect the reaction.
  • a solvent that does not adversely affect the reaction.
  • the solvent used include chloroform, dichloromethane, a mixed solution of benzene and water, acetonitrile, ethylalcohol, and the like.
  • the reaction temperature is not specifically limited and is generally carried out at reduced or elevated temperature, preferably at room temperature. Desired products can be isolated and purified by general methods, for example, column chromatography or recrystallization.
  • Method 2 for preparing a compound of the formula (lb) first, a compound of the formula (H) is reacted with a compound of the formula (Vi ⁇ ) to produce a compound (IX). The reaction is generally carried out in the same manner as in the amidation for converting the compound (H) into the compound (IV) as described in Method 1.
  • the compound of the formula (IX) is subjected to nitration to produce the compound of the formula (lb).
  • This reaction is generally carried out in the same manner as in the nitration for converting the compound of the formula (IV) into the compound of the formula (la) as described in Method 1.
  • the compound of the formula (lb) can be synthesized by reacting the compound of the formula (IT) with a compound of the formula (X) to produce a compound (XI), followed by converting the same into the compound (lb).
  • the conditions of reacting the compound of the formula (II) with the compound of the formula (X) are the same as those for amidation like in the reaction between the compound of the formula (H) and the compound of the formula (IV) as described in Method 1.
  • the reaction is generally carried out under nitrating conditions of an alcoholic compound, that is, under the same conditions as those of converting the compound of the formula (VI) into the compound of the formula (la) as described in Method 1.
  • the compound of the formula (lb) can also be synthesized by performing nitration the compound of the formula (XU) prepared by converting hydrogen of alcohol in the compound of the formula (XI) into a leaving group such as methane sulfonyl, -toluene sulfonyl or triflate.
  • the reaction is generally carried out under nitrating conditions, that is, under the same conditions of converting the compound of the formula (VIT) into the compound of the formula (la) as described in Method 1.
  • Desired products can be isolated and purified by general methods, for example, column chromatography or recrystallization.
  • Method 3 for preparing a compound of the formula (Ic) first, a compound of the formula (HI) is reacted with a compound of the formula (X1TI) to produce a compound (XIV).
  • the base examples include pyridine, 4-dimethylaminopyridine, triethylamine, diethylisopropylamine, dimethylphenylamine, 2,6-lutidine, or sodium hydride (NaH), cesium carbonate, and a phase transfer catalyst such as sodium hydroxide or benzyltriethylammonium chloride.
  • the reaction can be advantageously carried out in the presence of a solvent that does not adversely affect the reaction. Examples of the solvent include dichloromethane, chloroform, tetrahydrofuran, diethylether, toluene, dimethylformamide, toluene, dimethylformamide, acetonitrile, and the like.
  • the reaction temperature is not specifically limited and is generally carried out at reduced or elevated temperature, preferably at room temperature.
  • the compound of the formula (XIV) synthesized in the first step is subjected to nitration to produce the compound of the formula (Ic). This reaction is generally carried out in the same manner as in the nitration of halogen for converting the compound of the formula (IV) into the compound of the formula (la) as described in Method 1.
  • the compound of the formula (Ic) can be synthesized by reacting the compound of the formula (Xm) with a compound of the formula (XV) to produce a compound (XVI), followed by converting the same into the compound (Ic).
  • the reaction is generally carried out under nitrating conditions of an alcoholic compound, that is, under the same conditions as those of converting the compound of the formula (VI) into the compound of the formula (la) as described in Method 1.
  • the compound of the formula (Ic) also can be synthesized by performing nitration the compound of the formula (XVH) prepared by converting hydrogen of alcohol in the compound (XVI) is converted into a leaving group such as methane sulfonyl, p-tohxene sulfonyl or triflate.
  • the reaction is generally carried out under nitrating conditions of an alcoholic compound, that is, under the same conditions of converting the compound of the formula (VH) into the compound of the formula (la) as described in Method 1.
  • the compound of the formula (Ic) ' can be synthesized by reacting the compound of the formula (XHT) with a compound of the formula (XVJJf) having a protecting group in alcohol to convert the same into a compound of the formula (XIX), followed by converting the compound of the formula (XIX) into the compound (XVI) by a deprotection.
  • the reaction is preferably carried out in the presence of a base, preferably pentafluorophenyl and pyridine, or ethylchloroformate and triethylamine.
  • a base preferably pentafluorophenyl and pyridine, or ethylchloroformate and triethylamine.
  • This reaction can also be carried out without using a solvent, preferably in the presence of a solvent that does not adversely affect the reaction. Examples of the solvent include dichloromethane, chloroform, tetrahydrofuran, diethylether, and the like.
  • the reaction temperature is not specifically limited and is generally carried out at reduced or elevated temperature, preferably at room temperature.
  • Desired products can be isolated and purified by general methods, for example, column chromatography or recrystallization.
  • Method 4 for preparing a compound of the formula (Id) first, a compound of the formula (XX) is reacted with a compound of the formula (VHI) to produce a compound (XXI). The reaction is generally carried out in the same manner as in the amidation for converting the compound (H) into the compound (IV) as described in Method 1.
  • the compound of the formula (XXI) synthesized in the first step is subjected to nitration to produce the compound of the formula (Id).
  • This reaction is generally carried out in the same manner as in the nitration of halogen for converting the compound of the formula (IV) into the compound of the formula (la) as described in Method 1.
  • the compound of the formula (Id) can be synthesized by reacting the compound of the formula (XX) with a compound of the formula (X) to produce a compound (XXH), followed by converting the same into the compound (Id).
  • the conditions of reacting the compound of the formula (XX) with the compound of the formula (X) are the same as those for amidation by which the compound of the formula (U) is converted into the compound of the formula (IV) as described in Method 1.
  • the reaction is generally carried out under nitrating conditions of an alcoholic compound, that is, under the same conditions as those of converting the compound of the formula (VI) into the compound of the formula (la) as described in Method 1.
  • the compound of the formula (Id) can also be synthesized by performing nitration the compound of the formula (XXIH) prepared by converting hydrogen of alcohol in the compound of the formula (XXH) into a leaving group such as methane sulfonyl, ?-toluene sulfonyl or triflate.
  • the reaction is generally carried out under nitrating conditions, that is, under the same conditions of converting the compound of the formula (VH) into the compound of the formula (la) as described in Method 1.
  • Desired products can be isolated and purified by general methods, for example, column chromatography or recrystallization.
  • Method 5 for preparing a compound of the formula (le) first, a compound of the formula (XHT) is reacted with a compound of the formula (VET) to produce a compound (XXIV).
  • the base include sodium hydride (NaH), cesum carbonate, silver carbonate, a phase transfer catalyst such as sodium or potassium hydroxide or benzyltriethylammonium chloride, or crown ether.
  • the reaction is preferably carried out in the presence of a solvent that does not adversely affect the reaction.
  • a solvent include dichloromethane, chloroform, tetrahydrofuran, diethylether, toluene, dimethylformamide, dimethyl sulfoxide, or benzene.
  • the reaction temperature is not specifically limited and is generally carried out at reduced or elevated temperature, preferably at cooling temperature or room temperature.
  • the compound of the formula (XXTV) synthesized in the first step is subjected to nitration to convert the same into a compound of the formula (le).
  • the reaction is generally carried out in the same manner as in the nitration of halogen for converting the compound (IV) into the compound (la) as described in Method 1.
  • the compound of the formula (le) can be synthesized by reacting the compound of the formula (XHI) with a compound of the formula (X) to produce a compound (XXV), followed by converting the same into the compound (le).
  • the conditions of reacting the compound of the formula (XHT) with the compound of the formula (X) are the same as those for the reaction between the compound of the formula (XHT) and the compound of the formula (VIH).
  • the reaction is generally carried out under nitrating conditions of an alcoholic compound, preferably under the same conditions for the reaction for converting the compound of the formula (VI) into the compound of the formula (la) as described in Method 1.
  • the compound of the formula (le) can also be synthesized by performing nitration a compound of the formula (XXVI) prepared by converting hydrogen in alcohol in the compound of the formula (XXV) into a leaving group such as methane sulfonyl, /?-toluene sulfonyl or triflate.
  • a compound of the formula (XXVI) prepared by converting hydrogen in alcohol in the compound of the formula (XXV) into a leaving group such as methane sulfonyl, /?-toluene sulfonyl or triflate.
  • the reaction is generally carried out under nitrating conditions, preferably under the same conditions of converting the compound of the formula (VH) into the compound of the formula (la) as described in Method 1.
  • Desired products can be isolated and purified by general methods, for example, column chromatography or recrystallization.
  • the pharmaceutical composition according to the present invention including the colchicine derivative of the formula (I) and its pharmaceutically acceptable salt as effective components can be used for gout treatment agents, anticancer agents, anti-proliferous agents, anti-inflammatory agents, immunosuppressive agents and muscle relaxing agents.
  • the pharmaceutical composition according to the present invention can be prepared in various parenterally or orally administrable formulations.
  • Typical examples of formations for parenteral administration preferably include in the form of an isotonic aqueous saline solution or suspension for injection.
  • formulations for oral administration include tablets, capsules and the like, which may further contain a diluent (e.g.: lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine) or a lubricant (e.g.: silica, talc, stearic acid and its magnesium or potassium salt, and/or polyethylene glycol) in addition to effective components.
  • a diluent e.g.: lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine
  • a lubricant e.g.: silica, talc, stearic
  • Tablets can further be prepared with a binder such as magnesium aluminum silicate, starch paste, gelatins, tragacanth, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidine.
  • a binder such as magnesium aluminum silicate, starch paste, gelatins, tragacanth, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidine.
  • an disintegrator such as starch, agar, and alginate or sodium salts thereof, boiling mixtures and/or absorbents, a coloring agent, a flavoring agent, or a sweetener.
  • the formulations can be prepared by general techniques of mixing, granulation or coating.
  • the pharmaceutical composition according to the present invention is sterilized and/or contains additives such as an antiseptic, a stabilizer, a hydrator or emulsifier, osmosis adjusting salts and/or a buffering agent, and other therapeutically effective materials.
  • additives such as an antiseptic, a stabilizer, a hydrator or emulsifier, osmosis adjusting salts and/or a buffering agent, and other therapeutically effective materials.
  • the colchicine derivative of the formula (I) and its pharmaceutically acceptable salt can be administered to mammals including humans through parenteral or oral routes in an amount of 1 to 200 mg/kg (body weight) once or several times per a day.
  • the mixture was extracted with chloroform, dried over anhydrous magnesium sulfate and filtered, and then concentrated under reduced pressure to obtain 4-iodo-N-(l,2,3,10-tetramethoxy-9-oxo-5,6,7,9- tertahydro-benzo[a]heptarene-7-yl)-butylamide (intermediate compound 1) (the first step reaction).
  • the concentrated intermediate compound 1 and 1ml of acetonitrile were placed into a 15ml flask and 0.15g (0.87mmol) of silver nitrate was added thereto. After stirring for 12 hours, the mixture was filtered. The solvent was removed under reduced pressure.
  • the compound 13 obtained from the Example 13 was dissolved using 5ml of acetone. 0.072g (0.48mmol) of the sodium iodide was added dropwise thereto. After stirring at 40 ⁇ 50 ° C for 24 hours, the mixture was extracted with chloroform, dried over anhydrous magnesium sulfate and filtered. The mixture was concentrated under reduced pressure to obtain N-(7-acetylamino-l,2,3-trimethoxy-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene- 10-yl)-4-iodidmethyl benzamide (intermediate compound 7).
  • Table 1 shows the list of the compounds synthesized in accordance with the method of the present invention in addition to the compounds of the above Examples Table 1A
  • MCF-7/DOX (adriamycin resistant cell strain), MRS-SA (human uterine sarcoma), MES-SA/DX5 (adriamycin resistant cell strain), A 549 (human non-small cell lung), SKOV-3 (human ovarian), SKMEL-2 (human melanoma), XF-498 (human CNS), HCT-15 (human colon) were incubated at 37 ° C in the presence of 5% CO 2 using a DMEM culture medium. The respective cells were seeded into each well of 96-well plates at a concentration of 2xl0 3 ⁇ 5> ⁇ 10 3 cells/well.
  • the colchicine derivative of the present invention exhibited a higher anticancer effect even at a low concentration of 0.02 to 773 nM than the conventional colchicine and taxol as shown in Tables 2 and 3.
  • Example B Mixed lymphocyte reaction (MLR) tests MLR tests were carried out to determine the immunosuppressive effect of an immunosuppressive candidate material.
  • a responding cell BALB/c mouse spleen cell
  • a stimulating cell DBA/2 mouse spleen cell
  • the cells grew little.
  • the proliferated cells were treated with the immunosuppressive candidate material for measurement of the proliferation inhibitory extent.
  • Responding cells BALB/c mouse spleen cells
  • stimulating cells DBA/2 mouse spleen cells
  • cyclosporin A positive control group
  • colchicines colchicine derivatives of the present invention
  • the colchicine derivatives according to the present invention that is, the compounds 6, 9, 10, 11 and 12, suppressed growth of cells even at concentrations as low as 100 to 1000 nm and had a good immunosuppressive effect.
  • Example C Immunosuppressive effect test using BALB/c mouse spleen cells
  • This test was carried out to determine the immunosuppressive effect of an immunosuppressive candidate material by checking anti-proliferation of T cells and B cells.
  • responding cells BALB/c mouse spleen cells
  • lipopolysaccharide LPS
  • ConA concanvalin A
  • Responding cells (BALB/c mouse spleen cells) were seeded into each well of 96-well plates at a concentration of 2*10 5 cells/well. Then, 20 ⁇ g/ml lipopolysaccharide, cyclosporin A (positive control group), and inventive colchicine derivatives (compounds 6, 10 and 12) were simultaneously treated, and 2 ⁇ g/ml of ConA, cyclosporin A (positive control sample), and inventive colchicine derivatives (compounds 6, 10 and 12) were simultaneously treated. After culturing for 72 hours at a CO 2 incubator, a lO ⁇ A MTS solution was added to each well, followed by further culturing for 2 to 4 hours and measuring OD at 490 nm using ELISA. The measurement results are shown in FIGS. 2 and 3.
  • the colchicine derivatives according to the present invention suppressed proliferation of the B and T cells induced by LPS and ConA, respectively, in a concentration-dependent manner.
  • the immunosuppressive effect of the colchicine derivatives according to the present invention was superior to that of cyclosporin A, a conventional immunosuppressive agent.
  • the compounds 6 and 12 exhibited a remarkable anti-proliferous effect on B and T cells (FIG. 3).
  • Example D Toxicity assay
  • the colchicine derivatives according to the present invention were administered to ICR mice of 4-5 weeks old, weighing 18 to 20g, for acute (intravenous administration) toxicity assay and oral administration toxicity assay, as demonstrated in Tables 5 and 7.
  • the same assays were carried out using colchicine as a control group (see Tables 4 and 6).
  • This assay is carried out to test mutagenicity of chemical substances using histidine auxotrophic strains that are one of Salmonella Typhimurium mutants.
  • artificially induced mutants histidine auxotrophic mutants
  • histidine-free culture medium When various mutagens are added to the culture medium, only revertants generated by reverse mutation survive. Thus generated colonies and spontaneously induced revertants are compared for detection of mutation.
  • Ames test was carried out using WP2 uvrA strains (tryptophan auxotrophic strains) of TA100, TA1535, TA98, TA1537 and Escherichia coli.
  • Mutagens used as positive control samples were 0.5 g/plate of sodium azide, 0.5 g/plate of 4NQO (4-nitroquinolin-l -oxide), 50 g/plate of 9-AA (9-aminoacridine).
  • Colchicine and colchicine derivative (Compound 6) according to the present invention were treated at amounts of 0, 317.5, 625, 1250, 2500 and 5000 g/plate, and incubated in the presence (+) and absence (-) of microsomal polysubsfrate oxygenases (S-9 mixture) at 37 ° C for 48 hours. After incubation, the number of revertant colonies was counted. 3 plates were prepared for each test and the average was calculated. The results are demonstrated in Tables 8 and 9.
  • the colchicine derivatives according to the present invention induced noticeably fewer revertant colonies than the positive control groups, implying no significant hazard as mutagens.
  • novel colchicine derivative of the formula (I) according to the present invention or pharmaceutically acceptable salts thereof are superior to conventional colchicine in view of anticancer, anti-proliferous and immunosuppressive effects, and have less likelihood of toxicity and less hazard as mutagen than conventional colchicine.

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Abstract

Colchicine derivatives represented by the formula (I) with a halogen or nitric ester group, or pharmaceutically acceptable salts thereof, are described. Pharmaceutical compositions containing the same as effective components are also described. The colchicine derivatives were found to have anticancer, anti-proliferous and immunosuppressive function. Methods for preparing the colchicines derivatives are also provided.

Description

A NOVEL ALKALOID DERIVATIVE AND A PHARMACEUTICAL COMPOSITION CONTAINING THE SAME
Technical Field The present invention relates to a novel alkaloid derivative and a pharmaceutical composition containing the same. More particularly, the present invention relates to a novel colchicine derivative having anticancer, anti-proliferous and anti-inflammatory effects and immunosuppressive and muscle relaxing functions, and pharmaceutically acceptable salts thereof, a pharmaceutical composition containing the same as an effective component, and methods for preparing the same.
Background Art
Colchicine is a pseudo-alkaloid widely used for treatment of gout and is used only for short-term therapeutic treatment due to its toxicity. However, colchicine has been reported to exhibit a very fast and specific therapeutic effect on gout, as described in the Alkaloids, 1991, vol. 41, 125-176, U.S. patent No. 4,533,675, and so on.
During cell division, colchicine inhibits formation of mitotic spindle, thereby suppresses cell division, leading to activation of anticancer and anti-proliferous effects. Also, continuous research into colchicine applications has been carried out and a variety of colchicine derivatives have been synthesized up to now, as described in U.S. patent No. 3,222,253, U.S. patent application serial No. 00/608073A, WO
91/02084, and so on. Among them, only demecolcine has been used for treatment of leukemia. There is also a report that colchicine can be used for treatment of psoriasis or rheumatoid arthritis and has an amyloidosis inhibitory effect and an anti-inflammatory effect (Arch. Dermatol. 1982, Vol. 118, July, pp 453-457). Also, thiocolchicoside, which is one of colchicine derivatives, is widely used for treatment of skeletal muscle contracture and inflammation.
Disclosure of the Invention A first feature of the present invention is to provide a novel colchicine derivative having anticancer, anti-proliferous and anti-inflammatory effects and immunosuppressive and muscle-relaxing functions, and pharmaceutically acceptable salts thereof. A second feature of the present invention is to provide methods for preparing the colchicine derivatives.
A third feature of the present invention is to provide a pharmaceutical composition containing the colchicine derivative and pharmaceutically acceptable salts thereof as an effective component. In an aspect of the present invention, it is provided colchicine derivatives represented by the formula (I) and pharmaceutically acceptable salts thereof:
wherein when Ri is N(R6)C(X])-A, X2C(Xχ)-A, N(Re)-A, N(A)2 or X2-A, R2 is X3R7 or
N(R )2, R3 and R-t are independently hydrogen or a methyl group, R5 is hydrogen, a methyl group or CH2X4R7, wherein R^ and R7 are independently hydrogen or a lower alkyl, and X, Xi, X2, X and X4 are independently O or S; when R2 is N(Re)C(Xι)-A, X2C(X -A, N(Re)-A, N(A)2- or X2-A, R, is N(R6)COCH3, N(Re)COCF3, or NHC(O)OR8, R3 and R4 are independently hydrogen or a methyl group, R5 is hydrogen, a methyl group or CH2X4R , wherein Rβ and R7 are independently hydrogen or a lower alkyl, R8 is a lower alkyl, alkenyl, or substituted or unsubstituted aryl, and X, X\, X2 and X4 are independently O or S; when R and R4 are independently C(Xι)-A or -A, Ri is N(R )COCH3, N(R6)COCF3, or NHC(O)OR8, R2 is X3R7 or N(R7)2, R5 is hydrogen, a methyl or
CH2X R7, wherein Rό and R7 are independently hydrogen or a lower alkyl, R8 is a lower alkyl, alkenyl, or substituted or unsubstituted aryl, and X, X\, X3 or are independently O or S; when R5 is CH2X2C(X,)-A, R, is N(R6)COCH3, N(R6)COCF3 or NHC(O)OR8, R2 is X3R7 or N(R7)2, R3 and ^ are independently hydrogen or a methyl group, wherein R$ and R7 are independently hydrogen or a lower alkyl, R8 is a lower alkyl, alkenyl, or substituted or unsubstituted aryl, and X, Xι,X2 and X are independently O or S, wherein A is represented by the formula (a), (b), (c), (d), (e), (f), (g), (h), (i) or
0):
— Y,-Hal (a) — Y,-ON02 (b)
-(CH-CH2-0)n,-Hal (c) -(CH-CH2-0)n,-N02 (d)
wherein Yj is a Ci to Cι0 straight chain or branched alkyl, preferably, a C2 to C5 straight chain or branched alkyl or a substituted C5 to C7 cycloalkyl; Hal is halogen, for example, F, Cl, Br or I, R9 is hydrogen or a lower alkyl; ni is an integer from 1 to 6, preferably from 2 to 4; n2 and n3 are independently an integer from 1 to 5, preferably from 1 to 3; is an integer from 0 to 3; and n5 is an integer from 1 to 6.
In another aspect of the present invention, it is provided a pharmaceutical composition containing the colchicine derivatives and pharmaceutically acceptable salts thereof as effective components, the pharmaceutical composition having anticancer, anti-proliferous and anti-inflammatory effects and immunosuppressive and muscle-relaxing functions. In still another aspect of the present invention, it is provided methods for preparing the colchicine derivative represented by the formula (I) according to the reaction schemes 1, 2, 3, 4 and 5. In the reaction schemes 1 to 5, the colchicine derivatives of the formula (I) are represented as the formula (la), (lb), (Ic), (Id), or (Ie):
Reaction scheme 1 (Method 1)
Reaction scheme 2 (Method 2)
Reaction scheme 3 (Method 3)
XVI XVH
XIX
Reaction scheme 4 (Method 4)
Reaction scheme 5 (Method 5)
XXV XXVI
wherein B in the formulas (la), (lb), (Ic), (Id) and (le) is represented by the formula Cl or C2, and C in the formulas is represented by the formula Cl, C2, C3, C4 or C5: wherein Ri through R5 and X are defined as in the compound of the formula
(i).
Rό is hydrogen or a lower alkyl; X\ and X2 are independently O or S; Hal, Hah, and Hal2 may be the same or different halogens; L is a leaving group selected from methanesulfonyl, ?-toluenesulfonyl or triflate; P represents a general hydroxy-protecting group including methoxymethyl or t-butyldimethylsilyl; R10 is hydrogen or a lower alcohol of to C3; and Y is represented by the general formula (a'Mb'Mc'Md^ or ^):
— Y,- (a1)
-(CH-CH^OJn, (b1)
— (CH2)n2-CH-(CH2)n3-CH3 (c1)
wherein Yi represents a Ci to Cι0 straight chain or branched alkyl, preferably, a C2 to C5 straight chain or branched alkyl, or a substituted C5 to C7 cycloalkyl, R8 represents hydrogen or a lower alkyl, ni is an integer from 1 to 6, preferably an integer from 2 to 4, n2 and n3 are independently an integer from 1 to 5, preferably an integer from 1 to 3, r is an integer 0 to 3, and n5 is an integer from 1 to 6.
Brief Description of the Drawings
FIG. 1 is a graph showing the immonosuppressive effect of a colchicine derivative tested by mixed lymphocyte reaction (MLR);
FIG. 2 is a graph showing the immonosuppressive effect of cyclosphorine A as a positive control sample in an immunosuppressive effect test using a BALB/c mouse spleen; and
FIG. 3 is a graph showing the immonosuppressive effect of a colchicine derivative according to the present invention in an immunosuppressive effect test using a BALB/c mouse spleen.
Best mode for carrying out the Invention
The present invention will now be described in more detail.
Throughout the specification, a lower alkyl represents a saturated Ci to C6, preferably, Cx to C4 straight chain or branched hydrocarbon.
Preferred examples of the colchicine derivative of the formula (1) and its pharmaceutically acceptable salt according to the present invention include:
4-chloro-N-(l,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl)- butylamide;
4-chloromethyl-N-(l,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-benzamide; 3-chloromethyl-N-(l,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-benzamide;
4-iodo-N-(l,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl)- butylamide;
4-nitrooxy-N-(l,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-7- yl)-butylamide;
4-iodomethyl-N-(l ,2,3, 10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene- 7-yl)-benzamide;
4-nitrooxymethyl-N-(l ,2,3, 10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-benzamide;
3-iodomethyl-N-(l,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene- 7-yl)-benzamide;
3-nitrooxymethyl-N-(l ,2,3, 10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-benzamide;
4-chloro-N-( 1 ,2, 3 -trimethoxy- 10-methylsulfanyl-9-oxo-5 ,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-butylamide; 4-chloromethyl-N-(l,2,3-trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-benzamide;
3-chloromethyl-N-(l,2,3-trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-benzamide;
4-iodo-N-(l,2,3-trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-butylamide;
4-nitrooxy-N-(l,2,3-trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-butylamide;
4-iodomethyl-N-( 1 ,2,3-trimethoxy- 10-methylsulfanyl-9-oxo-5 ,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-benzamide; 4-nitrooxymethyl-N-(l,2,3-trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-benzamide;
3-iodomethyl-N-(l,2,3-trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-benzamide;
3-nitrooxymethyl-N-(l,2,3-trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-benzamide;
N-(7-acetylamino-l,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-10- yl)-4-chloromethyl-benzamide;
N-(7-acetylamino-l,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-10- yl)-4-iodomethyl-benzamide; N-(7-acetylamino- 1 ,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene- 10- yl)-4-nitrooxymethyl-benzamide; 4-chloromethyl-benzoic acid 7-acetylamino- 1, 2,3, 10-tetramethoxy-9-oxo-5, 6,7,9- tetrahydro-benzo[a]heptarene-7-yl-rnethyl ester;
4-chloromethyl-benzoic acid 7-acetylamino- 1 ,2,3-trimethoxy- 10-methylsulfanyl-
9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl-methyl ester; 4-chloro-butyric acid 7-acetylamino- 1,2,3, 10-tetramethoxy-9-oxo-5, 6,7,9-tetrahydro- benzo[a]heptarene-7-yl-methyl ester;
4-chloro-butyric acid 7-acetylamino- 1 ,2,3-trimethoxy- 10-methylsulfanyl-9-oxo-
5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl-methyl ester;
4-nitrooxymethyl-benzoic acid 7-acetylamino- 1,2,3, 10-tetramethoxy-9-oxo-5, 6,7,9- tetrahydro-benzo[a]heptarene-4-yl-methyl ester;
4-iodomethyl-butyric acid 7-acetylamino- 1 ,2,3-trimethoxy- 10-methylsulfanyl-9-oxo-
5,6,7,9-tetrahydro-benzo[a]heptarene-4-yl-methyl ester;
4-nitrooxymethyl-butyric acid 7-acetylamino- 1 ,2,3-trimethoxy- 10-methylsulfanyl-
9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-4-yl-methyl ester; 4-iodomethyl-benzoic acid 7-acetylamino- 1 ,2,3-trimethoxy- 10-methylsulfanyl-9- oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-4-yl-methyl ester;
4-nitrooxymethyl-benzoic acid 7-acetylamino- 1 ,2,3-trimethoxy- 10-methylsulfanyl-9- oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-4-yl-methyl ester;
(-)-3-chloromethyl-benzoic acid 1,2,3, 10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl ester;
(+)-3-chloromethyl-benzoic acid 1,2,3, 10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl ester;
(-)-3-iodomethyl-benzoic acid 1 ,2,3, 10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl ester; (+)-3-iodomethyl-benzoic acid 1, 2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl ester;
(-)-3-nitrooxymethyl-benzoic acid 1 ,2,3, 10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl ester;
(+)-3-nitrooxymethyl-benzoic acid 1, 2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl ester;
4-chloro-butyric acid 7-acetylamino- 1 ,2, 10-trimethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-3-yl ester;
4-chloromethyl-benzoic acid 7-acetylamino-l,2,10-trimethoxy-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-3-yl ester;
3-chloromethyl-benzoic acid 7-acetylamino- 1,2,10-trimethoxy-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-3-yl ester;
4-nitrooxy-butyric acid 7-acetylamino- 1,2,10-trimethoxy-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-3-yl ester;
4-nitrooxymethyl-benzoic acid 7-acetylamino- 1, 2, 10-trimethoxy-9-oxo-5, 6,7,9- tetrahydro-benzo[a]heptarene-3-yl ester; 3-nitrooxymethyl-benzoic acid 7-acetylamino-l,2,10-trimethoxy-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-3-yl ester;
4-chloro-N-methyl-N-(l,2,3-trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-butylamide;
4-nitrooxymethyl-N-methyl-N-(l,2,3-trimethoxy-10-methylsulfanyl- 9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-7-yl)-benzamide;
3-nitrooxymethyl-N-methyl-N-(l,2,3-trimethoxy-10-methylsulfanyl- 9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-7-yl)-benzamide;
(-)-3-chloromethyl-benzoic acid 1 ,2,3-trimethoxy- 10-methylsulfenyl-9-oxo-5 ,6,7,9- tetrahydro-benzo[a]heptarene-7-yl ester; (+)-3-chloromethyl-benzoic acid l,2,3-trimethoxy-10-methylsulfenyl-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-7-yl ester;
(-)-3-nitrooxymethyl-benzoic acid 1 ,2,3-trimethoxy-10-methylsulfenyl-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-7-yl ester;
(+)-3-nitrooxymethyl-benzoic acid 1 ,2,3-trimethoxy- 10-methylsulfenyl-9-oxo-5, 6,7,9- tetrahydro-benzo[a]heptarene-7-yl ester;
4-chloro-N-methyl-N-(l ,2,3, 10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl) butylamide;
4-chloromethyl-N-methyl-N-(l ,2,3, 10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl) benzamide; N-methyl-4-nitrooxy-N-( 1,2,3, 10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl) butylamide; N-methyl-4-nitrooxymethyl-N-( 1 ,2,3 , 10-tetramethoxy-9-oxo-5 ,6,7,9-tetrahydro- benzo[a]heptarene-7-yl) benzamide;
4-nitrooxy-butyric acid 7-acetylamino-l,2,3,10-tetramethoxy-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-4-yl methyl ester; 4-chloro-N-(l,2,3-trimethoxy-4-methoxymethyl-10-methylsulfanyl-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-7-yl)-butylamide;
4-chloromethyl-N-(l,2,3-trimethoxy-4-methoxymethyl-10-methylsulfanyl-9-oxo-
5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl)-benzamide;
4-nitrooxy-N-( 1 ,2,3 -trimethoxy-4-methoxymethyl- 10-methylsulfanyl-9-oxo-5 ,6,7,9- tetrahydro-benzo[a]heptarene-7-yl)-butylamide;
4-nitrooxymethyl-N-( 1 ,2,3 -trimethoxy-4-methoxymethyl- 10-methylsulfanyl-9-oxo-
5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl)-benzamide;
N-(7-acetylamino-l,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-10- yl)-3-chloromethyl-benzamide; N-(7-acetylamino- 1 ,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene- 10- yl)-3-nitrooxymethyl-benzamide;
N-(7-acetylamino- 1 ,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene- 10- yl-4-chloro-butylamide;
4-chloromethyl-N-(10-dimethylamino-l,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-benzamide;
N-(10-dimethylamino-l,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-7
-yl)-4-nitrooxymethyl-benzamide;
3-chloromethyl-N-(10-dimethylamino-l,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-benzamide; N-(10-dimethylamino-l,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-7
-yl)-3-nitrooxymethyl-benzamide;
4-chloro-N-(10-dimethylamino-l,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-butyrilamide; and
N-(10-dimethylamino-l,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-7 -yl)-4-nitrooxy-butyrilamide. Examples of the pharmaceutically acceptable salt of the colchicine derivative of the formula (I) include, but are not limited to, salts with inorganic bases such as sodium, potassium, magnesium or calcium and salts with organic bases such as ammonium, lysine, ethanolamine, N,N'-dibenzylethylenediamine and angelic acid Preparation methods of the invention will now be described in more detail.
Method 1
In Method 1 for preparing a compound of the formula (la), first, a compound of the formula (H) is reacted with a compound of the formula (III) to produce a compound (IN). The reaction can be carried out without using a base, but is generally carried out in the presence of a base used for amidation. Preferred examples of the base include pyridine, triethylamine, diethylisopropylamine and dimethylphenylamine, and a phase transfer catalyst such as sodium hydrocarbonate or benzyltriethylammonium chloride. The reaction can be carried out without using a solvent, but is advantageously carried out in the presence of a solvent that does not adversely affect the reaction. Examples of the solvent used include dichloromethane, chloroform, tetrahydrofuran, diethylether, toluene and dimethylformamide. The reaction temperature is not specifically limited and is generally carried out at reduced or elevated temperature, preferably at room temperature.
Second, the compound of the formula (IV) is subjected to nitration to convert it into a compound of the formula (la). Compounds capable of nitrating halogen are generally used for the reaction, and examples of such compounds include AgNO3 and t-butylammonium nitrate (B114NNO3). The reaction is preferably carried out in the presence of a solvent that does not adversely affect the reaction, and examples of such solvent include chloroform, acetonitrile, a mixed solution of acetonitrile and water, dichloromethane, and the like. The reaction temperature is not specifically limited and is generally carried out at reduced or elevated temperature, preferably at room temperature.
Alternatively, the compound of the formula (la) can be synthesized by reacting the compound of the formula (U) with a compound of the formula (V) to produce a compound (VI), followed by converting it into the compound (la). The conditions for reacting the compound of the formula (H) with the compound of the formula (V) are the same as those for amidation like in the reaction between the compound of the formula (H) and the compound of the formula (III). In order to convert the compound (VI) into the compound of the formula (la), the reaction is generally carried out under nitrating conditions of alcohol. Preferably, nitric acid and sulfuric acid, dinitrogen pentaoxide (N2O5) and aluminum chloride HI, potassium nitrate and boron trifluoride (BF3), acetylnitrate, etc., may be used, most preferably nitric acid and acetic anhydride (Ac2O) are used. The reaction is preferably carried out in the presence of a solvent that does not adversely affect the reaction, and examples of the solvent used include chloroform, dichloromethane, and the like. The reaction temperature is not specifically limited and is generally carried out at reduced or elevated temperature, preferably at room temperature.
The compound of the formula (la) can also be synthesized by performing nitration a compound of the formula (VH) prepared by converting hydrogen of alcohol in the compound of the formula (VI) into a leaving group such as methane sulfonyl, 7-toluene sulfonyl or triflate. In order to convert the compound (VH) into the compound of the formula (la), the reaction is generally carried out under nitrating conditions. Most preferably, t-butylammonium nitrate (Bu^NNOs), t-butylammonium nitrate (Bu4NNO3) and nitric acid, nitric acid and silver nitrate, or potassium nitrate is used. The reaction is preferably carried out in the presence of a solvent that does not adversely affect the reaction. Examples of the solvent used include chloroform, dichloromethane, a mixed solution of benzene and water, acetonitrile, ethylalcohol, and the like. The reaction temperature is not specifically limited and is generally carried out at reduced or elevated temperature, preferably at room temperature. Desired products can be isolated and purified by general methods, for example, column chromatography or recrystallization. Method 2
In Method 2 for preparing a compound of the formula (lb), first, a compound of the formula (H) is reacted with a compound of the formula (Viπ) to produce a compound (IX). The reaction is generally carried out in the same manner as in the amidation for converting the compound (H) into the compound (IV) as described in Method 1.
Second, the compound of the formula (IX) is subjected to nitration to produce the compound of the formula (lb). This reaction is generally carried out in the same manner as in the nitration for converting the compound of the formula (IV) into the compound of the formula (la) as described in Method 1.
Alternatively, the compound of the formula (lb) can be synthesized by reacting the compound of the formula (IT) with a compound of the formula (X) to produce a compound (XI), followed by converting the same into the compound (lb). The conditions of reacting the compound of the formula (II) with the compound of the formula (X) are the same as those for amidation like in the reaction between the compound of the formula (H) and the compound of the formula (IV) as described in Method 1. In order to convert the compound (XI) into the compound of the formula (lb), the reaction is generally carried out under nitrating conditions of an alcoholic compound, that is, under the same conditions as those of converting the compound of the formula (VI) into the compound of the formula (la) as described in Method 1.
The compound of the formula (lb) can also be synthesized by performing nitration the compound of the formula (XU) prepared by converting hydrogen of alcohol in the compound of the formula (XI) into a leaving group such as methane sulfonyl, -toluene sulfonyl or triflate. In order to convert the compound (XIT) into the compound of the formula (lb), the reaction is generally carried out under nitrating conditions, that is, under the same conditions of converting the compound of the formula (VIT) into the compound of the formula (la) as described in Method 1.
Desired products can be isolated and purified by general methods, for example, column chromatography or recrystallization. Method 3
In Method 3 for preparing a compound of the formula (Ic), first, a compound of the formula (HI) is reacted with a compound of the formula (X1TI) to produce a compound (XIV). The reaction is generally esterification between alcohol (X2=O) or thioalcohol (X2=S) and acyl or thioacyl halide, that is, the reaction is carried out in the presence of mckelacetylacetonate or zinc chloride or in the presence of a base that can be used for esterification. Examples of the base include pyridine, 4-dimethylaminopyridine, triethylamine, diethylisopropylamine, dimethylphenylamine, 2,6-lutidine, or sodium hydride (NaH), cesium carbonate, and a phase transfer catalyst such as sodium hydroxide or benzyltriethylammonium chloride. Also, the reaction can be advantageously carried out in the presence of a solvent that does not adversely affect the reaction. Examples of the solvent include dichloromethane, chloroform, tetrahydrofuran, diethylether, toluene, dimethylformamide, toluene, dimethylformamide, acetonitrile, and the like. The reaction temperature is not specifically limited and is generally carried out at reduced or elevated temperature, preferably at room temperature. Second, the compound of the formula (XIV) synthesized in the first step is subjected to nitration to produce the compound of the formula (Ic). This reaction is generally carried out in the same manner as in the nitration of halogen for converting the compound of the formula (IV) into the compound of the formula (la) as described in Method 1. Alternatively, the compound of the formula (Ic) can be synthesized by reacting the compound of the formula (Xm) with a compound of the formula (XV) to produce a compound (XVI), followed by converting the same into the compound (Ic). The reaction between the compound of the formula (XHI) and the compound of the formula (XV) is generally esterification between alcohol (X2=O) or thioalcohol (X2=S) and acyl or thioacyl halide, that is, the reaction is carried out under the same conditions as the reaction between the compound of the formula (XIH) and the compound of the formula (HI) as mentioned above. In order to convert the compound (XVI) into the compound of the formula (Ic), the reaction is generally carried out under nitrating conditions of an alcoholic compound, that is, under the same conditions as those of converting the compound of the formula (VI) into the compound of the formula (la) as described in Method 1.
The compound of the formula (Ic) also can be synthesized by performing nitration the compound of the formula (XVH) prepared by converting hydrogen of alcohol in the compound (XVI) is converted into a leaving group such as methane sulfonyl, p-tohxene sulfonyl or triflate. In order to convert the compound (XVH) into the compound of the formula (Ic), the reaction is generally carried out under nitrating conditions of an alcoholic compound, that is, under the same conditions of converting the compound of the formula (VH) into the compound of the formula (la) as described in Method 1.
Also, the compound of the formula (Ic)'can be synthesized by reacting the compound of the formula (XHT) with a compound of the formula (XVJJf) having a protecting group in alcohol to convert the same into a compound of the formula (XIX), followed by converting the compound of the formula (XIX) into the compound (XVI) by a deprotection. The conversion of the compound of the formula (XHT) into the compound of the formula (XIX) is generally esterification between alcohol (X2=O) or thioalcohol (X2=S) and carboxylic acid (Rιo=H) or carboxylic acid ester (Rι0=a lower alkyl of Ci to C3). When Rι0 is H, the reaction is preferably carried out in the presence of a base, preferably pentafluorophenyl and pyridine, or ethylchloroformate and triethylamine. This reaction can also be carried out without using a solvent, preferably in the presence of a solvent that does not adversely affect the reaction. Examples of the solvent include dichloromethane, chloroform, tetrahydrofuran, diethylether, and the like. The reaction temperature is not specifically limited and is generally carried out at reduced or elevated temperature, preferably at room temperature.
Desired products can be isolated and purified by general methods, for example, column chromatography or recrystallization. Method 4
In Method 4 for preparing a compound of the formula (Id), first, a compound of the formula (XX) is reacted with a compound of the formula (VHI) to produce a compound (XXI). The reaction is generally carried out in the same manner as in the amidation for converting the compound (H) into the compound (IV) as described in Method 1.
Second, the compound of the formula (XXI) synthesized in the first step is subjected to nitration to produce the compound of the formula (Id). This reaction is generally carried out in the same manner as in the nitration of halogen for converting the compound of the formula (IV) into the compound of the formula (la) as described in Method 1. Alternatively, the compound of the formula (Id) can be synthesized by reacting the compound of the formula (XX) with a compound of the formula (X) to produce a compound (XXH), followed by converting the same into the compound (Id). The conditions of reacting the compound of the formula (XX) with the compound of the formula (X) are the same as those for amidation by which the compound of the formula (U) is converted into the compound of the formula (IV) as described in Method 1. In order to convert the compound (XXII) into the compound of the formula (Id), the reaction is generally carried out under nitrating conditions of an alcoholic compound, that is, under the same conditions as those of converting the compound of the formula (VI) into the compound of the formula (la) as described in Method 1.
The compound of the formula (Id) can also be synthesized by performing nitration the compound of the formula (XXIH) prepared by converting hydrogen of alcohol in the compound of the formula (XXH) into a leaving group such as methane sulfonyl, ?-toluene sulfonyl or triflate. In order to convert the compound (XXIH) into the compound of the formula (Id), the reaction is generally carried out under nitrating conditions, that is, under the same conditions of converting the compound of the formula (VH) into the compound of the formula (la) as described in Method 1.
Desired products can be isolated and purified by general methods, for example, column chromatography or recrystallization. Method 5
In Method 5 for preparing a compound of the formula (le), first, a compound of the formula (XHT) is reacted with a compound of the formula (VET) to produce a compound (XXIV). The reaction is generally a reaction between alcohol (X2=O) or thioalcohol (X2=S) and alkylhalide to produce ether or thioether, preferably in the presence of a base that can be used for etherification. Examples of the base include sodium hydride (NaH), cecium carbonate, silver carbonate, a phase transfer catalyst such as sodium or potassium hydroxide or benzyltriethylammonium chloride, or crown ether. The reaction is preferably carried out in the presence of a solvent that does not adversely affect the reaction. Examples of the solvent include dichloromethane, chloroform, tetrahydrofuran, diethylether, toluene, dimethylformamide, dimethyl sulfoxide, or benzene. The reaction temperature is not specifically limited and is generally carried out at reduced or elevated temperature, preferably at cooling temperature or room temperature.
Second, the compound of the formula (XXTV) synthesized in the first step is subjected to nitration to convert the same into a compound of the formula (le). The reaction is generally carried out in the same manner as in the nitration of halogen for converting the compound (IV) into the compound (la) as described in Method 1.
Alternatively, the compound of the formula (le) can be synthesized by reacting the compound of the formula (XHI) with a compound of the formula (X) to produce a compound (XXV), followed by converting the same into the compound (le). The conditions of reacting the compound of the formula (XHT) with the compound of the formula (X) are the same as those for the reaction between the compound of the formula (XHT) and the compound of the formula (VIH). In order to convert the compound (XXV) into the compound of the formula (le), the reaction is generally carried out under nitrating conditions of an alcoholic compound, preferably under the same conditions for the reaction for converting the compound of the formula (VI) into the compound of the formula (la) as described in Method 1.
The compound of the formula (le) can also be synthesized by performing nitration a compound of the formula (XXVI) prepared by converting hydrogen in alcohol in the compound of the formula (XXV) into a leaving group such as methane sulfonyl, /?-toluene sulfonyl or triflate. In order to convert the compound (XXVI) into the compound of the formula (le), the reaction is generally carried out under nitrating conditions, preferably under the same conditions of converting the compound of the formula (VH) into the compound of the formula (la) as described in Method 1.
Desired products can be isolated and purified by general methods, for example, column chromatography or recrystallization.
As described above, the pharmaceutical composition according to the present invention including the colchicine derivative of the formula (I) and its pharmaceutically acceptable salt as effective components can be used for gout treatment agents, anticancer agents, anti-proliferous agents, anti-inflammatory agents, immunosuppressive agents and muscle relaxing agents.
The pharmaceutical composition according to the present invention can be prepared in various parenterally or orally administrable formulations. Typical examples of formations for parenteral administration preferably include in the form of an isotonic aqueous saline solution or suspension for injection. Examples of formulations for oral administration include tablets, capsules and the like, which may further contain a diluent (e.g.: lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine) or a lubricant (e.g.: silica, talc, stearic acid and its magnesium or potassium salt, and/or polyethylene glycol) in addition to effective components. Tablets can further be prepared with a binder such as magnesium aluminum silicate, starch paste, gelatins, tragacanth, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidine. In some cases, there may be further contained an disintegrator such as starch, agar, and alginate or sodium salts thereof, boiling mixtures and/or absorbents, a coloring agent, a flavoring agent, or a sweetener. The formulations can be prepared by general techniques of mixing, granulation or coating.
The pharmaceutical composition according to the present invention is sterilized and/or contains additives such as an antiseptic, a stabilizer, a hydrator or emulsifier, osmosis adjusting salts and/or a buffering agent, and other therapeutically effective materials. These preparations can be formulated in accordance with known methods usually employed in the formulation process.
As the effective components of the pharmaceutical composition of the present invention, the colchicine derivative of the formula (I) and its pharmaceutically acceptable salt can be administered to mammals including humans through parenteral or oral routes in an amount of 1 to 200 mg/kg (body weight) once or several times per a day.
Examples
The present invention will be further described by the following Examples, but the Examples do not limit the scope of the invention.
7-deacetylcolchicine used in Examples was synthesized according to the method of the reference EP 0 493 064 and Synthetic Communications 1997, 27(2), 293-296.
The preparation of the thiodeacetylcolchicine was carried out in accordance with the method of WO 9421598 and Bioorganic & Medicinal Chemistry, Vol 5, No. 12, pp 2277-2282(1997).
The preparation of N-(10-amino-l,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)acetamide was carried out in accordance with the method of WO 9421598.
The preparation of 4-hydroxymethylcolchicine and 4-hydroxy -methylthiocolchicine was carried out in accordance with the method of the Brevet Canadien 778369 and Justus Liebigs Ann. Chem. 662, 105-113 (1963)
The preparation of 7-hydroxy-l,2,3-trimethoxy-10-methylsulphenyl-6,7,- dihydro-5H-benzo[a]heptarene-9-on and colchicone was carried out in accordance with the method of the J. Med. Chem. Vol.40, 961-965 (1997). Example 1
Preparation of 4-chloro-N-(l,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro -benzo[a]heptarene-7-yl)-butylamide
O.lg (0.28mmol) of deacetylcolchicine was placed into a 10ml flask and 3.5ml of tetrahydrofuran was added thereto and then dissolved. To the mixture, 0.2m£ (1.40mmol) of triethylamine was added dropwise and 0.035m£ (0.31mmol) of 4-chlorobutyryl chloride was added slowly. The mixture was stirred for 2 hours, and then extracted with chloroform and dried over anhydrous magnesium sulfate and filtered. The solvent in the mixture was removed under reduced pressure. The concentrated product was purified by the column chromatography (ethylacetate: methanol = 85:15) to obtain the title product:
1H NMR (500 MHz, CDC13) δ : 1.87-1.91 (m, 1H), 2.00-2.04 (m, 2H), 2.29-2.54 (m, 5H), 3.50 (t, J= 4.3 Hz, 2H), 3.67 (s, 3H), 3.91 (s, 3H), 3.95 (s, 3H), 4.01 (s, 3H), 4.65-4.68 (m, IH), 6.54 (s, IH), 6.88 (d, J = 11.0 Hz,lH), 7.35 (d, J= 11.0 Hz, IH), 7.51 (d, J= 6.5 Hz, IH), 7.52 (s, IH)
Example 2
Preparation of 4-chloromethyl-N-(l,2,3,10-tetramethoxy-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-7-yl)-benzamide
0.12g of title compound 2 (yields: 79%, as a yellow solid) was obtained in accordance with the same method with Example 1 except that 3-chloromethylbenzoyl chloride was used instead of 4-chlorobutyryl chloride:
1H NMR (500 MHz, CDC13) δ :2.05-2.06 (m, IH), 2.36-2.42 (m, 2H), 2.52-2.52 (m, IH), 3.75 (s, 3H), 3.91 (s, 3H), 3.96 (s, 3H), 4.00 (s, 3H), 4.44 (s, 2H), 4.86-4.88 (m, IH), 6.55 (s, IH), 6.90 (d, J= 11.0 Hz, IH), 7.18 (d, J = 8.5 Hz, 2H), 7.39 (d, J = 11.0 Hz, IH), 7.69 (s, IH), 7.78 (d, J = 8.5 Hz, 2H), 8.40 (d, J= 6.5 Hz, IH)
Example 3
Preparation of 3-chloromethyl-N-(l,2,3,10-tetramethoxy-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-7-yl)-benzamide
0.13g of the title compound 3 (yields: 92%, as a yellow solid) was carried out in accordance with the same method with Example 1 except that 4-chloromethyl benzoyl chloride was used instead of 4-chlorobutyryl chloride:
Η NMR (500 MHz, CDC13) δ :2.22-2.26 (m, IH), 2.39-2.48 (m, 2H), 2.53-2.57 (m, IH), 3.75 (s, 3H), 3.95 (s, 3H), 3.97 (s, 3H), 4.01 (s, 3H), 4.90-4.95 (m, IH), 5.30 (d,12.0 Hz, IH), 5.36 (d,12.0 Hz, IH), 6.56 (s, IH), 6.93 (d, J= 11.0 Hz, IH), 7.12 (t, J= 7.5 Hz, IH), 7.30 (d, J= 7.5 Hz, IH), 7.41 (d, J = 11.0 Hz, IH), 7.56 (d, J = 7.5 Hz, IH), 7.69 (s, IH), 7.77 (s, IH), 8.44 (d, J= 6.5 Hz, IH)
Example 4
Preparation of 4-nitrooxy-N-(l ,2,3, 10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-butylamide
O.lg (0.22mmol) of the compound 1 obtained from the Example 1 was placed into a 10ml flask and dissolved by adding the 5 ml of acetone. 0.097g (0.065 mmol) of sodium iodide was added dropwise thereto, and then the mixture was stirred at temperature of 40-50 °C for 12 hours. The mixture was extracted with chloroform, dried over anhydrous magnesium sulfate and filtered, and then concentrated under reduced pressure to obtain 4-iodo-N-(l,2,3,10-tetramethoxy-9-oxo-5,6,7,9- tertahydro-benzo[a]heptarene-7-yl)-butylamide (intermediate compound 1) (the first step reaction). The concentrated intermediate compound 1 and 1ml of acetonitrile were placed into a 15ml flask and 0.15g (0.87mmol) of silver nitrate was added thereto. After stirring for 12 hours, the mixture was filtered. The solvent was removed under reduced pressure. Chloroform was added thereto and the mixture was dried over sodium sulfate and filtered under reduced pressure. The resulting compound was purified by the column chromatography (ethylacetate:methanol = 85:15) to obtain the title compound 4 as a yellow solid (53mg, yields of the second step: 50%) (the second step reaction).
1H NMR of the intermediate compound 1 :
1H NMR (500 MHz, CDC13) δ : 1.86-1.89 (m, IH), 2.02-2.07 (m, 2H), 2.18-2.25 (m, IH), 2.35-2.47 (m, 3H), 2.51-2.55 (m, IH), 3.55 (t, 2H), 3.65 (s, 3H), 3.90 (s, 3H), 3.94 (s, 3H), 3.99 (s, 3H), 4.61-4.66 (m, IH), 6.54 (s, IH), 6.82 (d, lH), 7.30 (d, lH), 7.44 (s, IH), 7.46 (d, IH)
Η NMR of the Compound 4:
Η NMR (500 MHz, CDC13) δ : 1.96-1.20 (m, 3H), 2.29-2.39 (m, 4H), 2.51-2.52 (m, IH), 3.66 (s, 3H), 3.91 (s, 3H), 3.95 (s, 3H), 4.01 (s, 3H), 4.40-4.44 (m, 2H), 4.65-4.67 (m, IH), 6.54 (s, IH), 6.89 (d, J = 11.0 Hz, IH), 7.36 (d, J= 11.0 Hz, IH), 7.51 (s, IH), 7.55 (d, J= 6.5 Hz, IH)
MS m/z (relative intensity): 977.349([2M+H]+, 17), 846.271(6), 527.12 (9), 489.165([M+H]+, 100), 358.170([M+H]+-CO(CH2)3ONO2, 48), 341.46(48)
Example 5
Preparation of 4-nitrooxymethyl-N-(l,2,3,10-tetramethoxy-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-7-yl)-benzamide
33mg of the title compound 5 (yields of the second step reaction: 45%, as a yellow solid) was prepared by the same method of Example 4 through 4-iodomethyl-N-(l,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetra-hydro-benzo[a] heptarene-7-yl)-bezamide (intermediate compound 2) except for using compound 2 obtained from the Example 2.
Η NMR of intermediate compound 2:
Η NMR (500 MHz, CDC13) δ :2.01-2.09 (m, IH), 2.31-2.45 (m, 2H), 2.51-2.55 (m, IH), 3.75 (s, 3H), 3.91 (s, 3H), 3.96 (s, 3H), 4.00 (s, 2H), 4.86-4.89 (m, IH), 6.54 (s, IH), 6.90 (d, IH), 7.19 (d, 2H), 7.38 (d, IH), 7.67 (s, IH), 7.71 (d, 2H), 8.28 (d, J= 6.5 Hz, IH)
1H NMR of compound 5:
1H NMR (500 MHz, CDC13) δ :2.02-2.09 (m, IH), 2.35-2.58 (m, 3H), 3.76 (s, 3H), 3.91 (s, 3H), 3.97 (s, 3H), 4.01 (s, 3H), 4.86-4.91 (m, IH), 5.30 (s, 2H), 6.56 (s, IH), 6.92 (d, J= 11.0 Hz, IH), 7.19 (d, J= 8.2 Hz, 2H), 7.41 (d, J= 11.0 Hz, IH), 7.70 (s, lH), 7.81 (d, J= 8.2 Hz, 2H), 8.36 (d, J= 6.5 Hz, IH)
Example 6
Preparation of 3-nitrooxymethyl-N-(l ,2,3, 10-tetramethoxy-9-oxo-5 ,6,7,9- tetrahydro-benzo[a]heptarene-7-yl)-benzamide
25mg of the title compound 6 (yields of the second step reaction: 42%, as a yellow solid) was prepared by the same method of Example 4 through 3-iodomethyl-N-(l,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetra-hydro-benzo[a] heptarene-7-yl)-benzamide (intermediate compound 3) except for using compound 3 obtained from the Example 3.
1H NMR of the intermediate compound 3:
1H NMR (500 MHz, CDC13) δ :2.23-2.31 (m, IH), 2.38-2.48 (m, 2H),
2.56-2.60 (m, IH), 3.74 (s, 3H), 3.92 (s, 3H), 3.97 (s, 3H), 3.99 (s, 3H), 4.23 (d, IH), 4.28 (d, IH), 4.88-4.95 (m, IH), 6.59 (s, IH), 7.03 (d, IH), 7.13 (t,
IH), 7.33 (d, IH), 7.45 (d, IH), 7.59 (d, IH), 7.70 (s, IH), 7.78 (s, IH),
8.44 (d, IH)
Η NMR of the compound 6:
1H NMR (500 MHz, CDC13) δ :2.23-2.31 (m, IH), 2.40-2.50 (m, 2H), 2.55-2.61 (m, IH), 3.76 (s, 3H), 3.92 (s, 3H), 3.98 (s, 3H), 4.02 (s, 3H),
4.89-4.94 (m, IH), 5.10 (d, J= 12.3 Hz, IH), 5.20 (d, J= 12.3 Hz, IH), 6.57 (s,
IH), 6.94 (d, J= 11.0 Hz, IH), 7.14 (t, J= 7.9 Hz, IH), 7.28 (d, J= 7.9 Hz, IH),
7.44 (d, J= 11.0 Hz, IH), 7.60 (d, J= 7.9 Hz, IH), 7.70 (s, IH), 7.81 (s, IH),
8.56 (d, J= 6.5 Hz, IH) Example 7
Preparation of 4-chlroro-N-(l ,2,3-trimethoxy- 10-methyl-sulfonly-9-oxo- 5,6,7,9-tetra-hydro-benzo[a]heptarene-7-yl)-butylamide.
0.13g of the title compound 7 (yields: 86%, as a yellow solid) was prepared by the same method of Example 1 except that thiodeacetylcolchicine was used instead of deacetylcolchicine.
Η NMR (500 MHz, CDC13) δ : 1.86-1.93 (m, IH), 2.00-2.10 (m, 2H),
2.23-2.31 (m, IH), 2.37-2.55 (m, 4H), 2.45 (s, 3H), 3.53 (td, J = 6.5, 1.5 Hz,
2H), 3.67 (s, 3H), 3.91 (s, 3H), 3.95 (s, 3H), 4.68-4.73 (m, IH), 6.54 (s, IH),
7.09 (d, J = 10.6 Hz, IH), 7.32 (d, J = 10.6 Hz, IH), 7.35 (d, J = 7.3 Hz, IH),
7.42 (s, IH)
Example 8
Preparation of 4-chloro-N-(l,2,3-trimethoxy-10-methylsulfonyl-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-7-yl)-benzamide
0.12 g of the title compound 8 (yields: 89%, as a yellow solid) was prepared by the same method of Example 1 except that thiodeacetylcolchicine and 3-chloromethylbenzoyl chloride were used instead of deacetylcochicine and 4-chlorobutyryl chloride, respectively.
Η NMR (500 MHz, CDC13) δ :2.09-2.15 (m, IH), 2.31-2.38 (m, IH),
2.41-2.48 (m, IH), 2.44 (s, 3H), 2.55-2.59 (m, IH), 3.75 (s, 3H), 3.91 (s, 3H),
3.97 (s, 3H), 4.47 (s, 2H), 4.90-4.95 (m, IH), 6.56 (s, IH), 7.10 (d, J = 10.6
Hz, IH), 7.25 (d, J= 8.2 Hz, 2H), 7.35 (d, J= 10.6 Hz, IH), 7.56 (s, IH), 7.84
(d, J= 8.2 Hz, 2H), 8.11 (d, J= 7.3 Hz, IH)
Example 9 Preparation of 3-chloromethyl-N-(l ,2,3-trimethoxy-l O-methylsulfonyl -9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl)-benzamide
0.15g of the title compound 9 (yields: 90%, as a yellow solid) was prepared by the same method of Example 1 except that thiodeacetylcolchicine and 4-chloromethylbenzoyl chloride were used instead of deacetylcolchicine and 4-chlorobutyryl chloride, respectively.
Η NMR (500 MHz, CDC13) δ :2.20-2.25 (m, IH), 2.33-2.49 (m, 2H), 2.45 (s, 3H), 2.56-2.60 (m, IH), 3.75 (s, 3H), 3.92 (s, 3H), 3.96 (s, 3H), 4.37 (d,11.7 Hz, IH), 4.41 (d,l 1.7 Hz, IH), 4.93-4.98 (m, IH), 6.57 (s, IH), 7.13 (d, J= 10.6 Hz, IH), 7.17 (t, J= 7.6 Hz, IH), 7.34 (d, J= 7.6 Hz, IH), 7.38 (d, J = 10.6 Hz, IH), 7.65 (s, IH), 7.67 (d, J= 7.6 Hz, IH), 7.77 (s, IH), 8.35 (d, J = 7.3 Hz, IH)
Example 10
Preparation of 4-nitrooxy-N-(l ,2,3-trimethoxy- 10-methylsulfonyl-9-oxo- 5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl)-butylamide
30mg of the title compound 10 (yields of second step reaction: 33%, as a yellow solid) was prepared by the same method of Example 4 through 4-iodo-N-(l,2,3-trimethoxy-10-methylsulfonyl-9-oxo-5,6,7,9-tetrahydro-benzo[a] heptarene-7-yl)-butylamide (intermediate compound 4) except for using compound 7 obtained from the Example 7.
Η NMR of intermediate compound 4:
1H NMR (500 MHz, CDC13) δ :1.92-1.98 (m, IH), 2.01-2.12 (m, 2H), 2.24-2.55 (m, 5H), 2.45 (s, 3H), 3.50-3.54 (m, 2H), 3.68 (s, 3H), 3.91 (s, 3H), 3.95 (s, 3H), 4.71-4.74 (m, IH), 6.55 (s, IH), 7.11 (d, IH), 7.34 (d, IH), 7.53 (s, IH), 7.98 (d, IH)
1H NMR of compound 10: 1H NMR (500 MHz, CDC13) δ :1.87-1.93 (m, IH), 1.96-2.06 (m, 2H),
2.24-2.55 (m, 5H), 2.45 (s, 3H), 3.67 (s, 3H), 3.91 (s, 3H), 3.95 (s, 3H), 4.42-4.47 (m, 2H), 4.70-4.75 (m, IH), 6.54 (s, IH), 7.11 (d, J = 10.6 Hz, IH), 7.34 (d, J= 10.6 Hz, IH), 7.49 (s, IH), 7.69 (d, J= 7.6 Hz, IH) Example 11 Preparation of 4-nitrooxymethyl-N-(l,2,3-trimethoxy-10-methylsulfonyl-9- oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl)-benzamide
75 mg of the title compound 11 (yields of second step reaction: 58%, as a yellow solid) was prepared by the same method of Example 4 through 4-iodomethyl-N-(l,2,3-trimethoxy-10-methylsulfonyl-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-benzamide (intermediate compound 5) except for using compound 8 obtained from the Example 8.
1H NMR of intermediate compound 5:
1H NMR (500 MHz, CDC13) δ :2.15-2.20 (m, IH), 2.28-2.35 (m, IH), 2.38-2.44 (m, IH), 2.44 (s, 3H), 2.53-2.56 (m, IH), 3.75 (s, 3H), 3.90 (s, 3H), 3.96 (s, 3H), 4.27 (s, 2H), 4.92-4.97 (m, IH), 6.56 (s, IH), 7.12 (d, IH), 7.17 (d, 2H), 7.36 (d, IH), 7.67 (s, lH), 7.80 (d, 2H), 8.67 (d, IH) 1H NMR of compound 11 :
1H NMR (500 MHz, CDC13) δ :2.14-2.20 (m, IH), 2.30-2.48 (m, 2H), 2.45 (s, 3H), 2.55-2.59 (m, IH), 3.76 (s, 3H), 3.91 (s, 3H), 3.97 (s, 3H),
4.93-4.98 (m, IH), 5.29 (s, 2H), 6.56 (s, IH), 7.13 (d, J = 10.6 Hz, IH), 7.20
(d, J= 8.5 Hz, 2H), 7.37 (d, J= 10.6 Hz, IH), 7.66 (s, IH), 7.90 (d, J= 8.5 Hz,
2H), 8.64 (d, J= 7.3 Hz, IH) Example 12
Preparation of 3-nitrooxymethyl-N-(l ,2,3-trimethoxy- 10-methylsulfonyl-9- oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl)-benzamide
70mg of the title compound 12 (yields of second step reaction: 64%, as a yellow solid) was prepared by the same method of Example 4 through 3-iodomethyl-N-(l,2,3-trimethoxy-10-methylsulfonyl-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-benzamide (intermediate compound 6) except for using compound 9 obtained from the Example 9. 1H NMR of intermediate compound 6 :
Η NMR (500 MHz, CDC13) δ :2.26-2.45 (m, 3H), 2.45 (s, 3H), 2.54-2.58 (m, IH), 3.75 (s, 3H), 3.91 (s, 3H), 3.97 (s, 3H), 4.16 (d, lH), 4.23 (d, IH), 4.95-5.00 (m, IH), 6.57 (s, IH), 7.05 (t, IH), 7.16 (d, IH), 7.26 (d, IH), 7.39 (d, lH), 7.60 (d, IH), 7.77 (s, IH), 7.80 (s, IH), 8.82 (d, IH) 1H NMR of compound 12:
1H NMR (500 MHz, CDC13) δ :2.31-2.47 (m, 3H), 2.45 (s, 3H),
2.57-2.61 (m, IH), 3.77 (s, 3H), 3.92 (s, 3H), 3.98 (s, 3H), 4.94-4.99 (m, IH),
5.10 (d,12.0 Hz, IH), 5.18 (d,12.0 Hz, IH), 6.57 (s, IH), 7.12 (t, J = 7.6 Hz,
IH), 7.18 (d, J = 10.6 Hz, IH), 7.25 (d, J = 7.6 Hz, IH), 7.42 (d, J = 10.6 Hz, IH), 7.69 (s, lH), 7.78 -7.79 (m, 2H), 9.02 (d, J= 7.3 Hz, IH)
Example 13
Preparation of N-(7-acetylamino-l,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene- 10-yl)-4-chloromethyl-benzamide
lg (26mmol) of N-(10-amino-l,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro benzo[a]heptarene-7-yl) acetamide was placed into a 25ml flask and dissolved with 10ml of trahydrofuran. 2.8ml (20 mmol) of triethylamine was added thereto dropwise, and then the mixture was stirred for 30 minutes. 0.55g (2.9mmol) of 4-chloromethylbenzoyl chloride was added thereto and dissolved with stirring at room temperature for 48 hours, extracted with chloroform, dried over anhydrous magnesium sulfate and then filtered. After concentrating under reduced pressure, the concentrated resulting compound was purified through the column chromatography (chloroform: methanol=9:l) to obtained 1.36g of the title compound 13 (yields: 98%, as a yellow solid).
Η NMR (500 MHz, CDC13) δ : 1.87(m, IH), 2.02(s, 3H), 2.39(m, IH), 2.41(m, IH), 2.53(m, IH), 3.67(s, 3H), 3.91 (s, 3H), 3.92 (s, 3H), 4.68 (s, 2H), 4.71 (m, IH), 6.54 (s, IH), 6.94(d, J=10.0Hz, IH), 7.54-7.67 (m, 4H), 8.00 (m, 2H), 9.21(d, J=10.9Hz, IH), 10.32(s, IH) Example 14
Preparation of N-(7-acetylamino-l,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene- 10-yl)-4-nitrooxymethyl-benzamide
The compound 13 obtained from the Example 13 was dissolved using 5ml of acetone. 0.072g (0.48mmol) of the sodium iodide was added dropwise thereto. After stirring at 40~50°C for 24 hours, the mixture was extracted with chloroform, dried over anhydrous magnesium sulfate and filtered. The mixture was concentrated under reduced pressure to obtain N-(7-acetylamino-l,2,3-trimethoxy-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene- 10-yl)-4-iodidmethyl benzamide (intermediate compound 7). 0.0205g (0.033mmol) of the concentrated intermediate compound 7 and 0.2ml of the acetonitrile were placed into a 25ml flask and 0.022g (0.13mmol) of silver nitrate was added thereto. After stirring for 24 hours, the mixture was filtered, and followed by concentrating under reduced pressure, and chloroform was added thereto. The resulting mixture was dried over sodium sulfate, filtered, and followed by concentrating under reduced pressure. 18mg of the title compound 14 (yields of second step reaction: 96%, as a yellow solid) was obtained after purification by column chromatography (chloroform : methanol = 9:1)
Η NMR of intermediate compound 7:
1H NMR (500 MHz, CDC13) δ : 1.85 (m, IH), 2.02 (s, 3H), 2.29 (m, IH),
2.40 (m, IH), 2.54 (m, IH), 3.66 (s, 3H), 3.91 (s, 3H), 3.95 (s, 3H), 4.50 (s, 2H), 4.69 (m, IH), 6.54 (s, IH), 6.85 (d, J= 6.75Hz, IH), 7.53 (m, 3H), 7.60 (s, IH), 7.93 (d, J=8.2Hz, 2H), 9.20 (d, J=10.9Hz, IH), 10.30 (s, IH) Η NMR of compound 14: Η NMR (500 MHz, CDC13) δ : 1.85 (m, IH), 2.02 (s, 3H), 2.2 9(m, IH),
2.41 (m, IH), 2.54 (m, IH), 3.66 (s, 3H), 3.91 (s, 3H), 3.94(s, 3H), 4.69 (m, IH), 5.52 (s, 2H), 6.54 (s, IH), 6.73 (d, J=6.8Hz, IH), 7.51-7.60 (m, 4H), 8.03 (m, 2H), 9.19 (d, J=l 0.9Hz, IH), 10.33 (s, IH) Example 15
Preparation of 4-chloromethyl-benzoic acid-7-acetylamino- 1,2,3, 10- tetramethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl-methyl ester
0.05g (0.12mmol) of 4-hydroxymethylcolchicine (N-(4-hydroxymethyl-
1,2,3, 10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]-heptarene-7-yl)-acetamide) was placed into a 10ml flask and 3.5ml of chloroform was added thereto to dissolve the compound. To the mixture, 0.13ml (0.096mmol) of triethylamine was added dropwise and then 0.088mg (0.48mmol) of 4-chloromethylbenzoyl chloride was added slowly. After stirring for 18 hours, the mixture was extracted with chloroform, dried and filtered over anhydrous magnesium sulfate, and the solvent was removed under reduced pressure. Concentrated resulting compounds was purified using the column chromatography (ethylacetate:methanol = 85:15) to obtain 0.034 g of the title compound 15 (yields: 50.2%) as a white solid.
Η NMR (500 MHz, CDC13) δ :1.85-1.87(m, IH), 1.99(s, 3H), 2.21-2.23(m, 2H), 2.92-2.94(m, IH), 3.67(s, 3H), 3.98(s, 3H), 4.00(s, 3H), 4.02(s, 3H), 4.61(s, 3H), 4.67-4.69(m, IH), 5.40(d, J=11.15Hz, IH), 5.46(d, J=11.15Hz, IH), 6.86 (d, J= 11.15Hz, IH), 7.30 (d, J= 10.85Hz, IH), 7.45(d, J=8.80Hz, 2H), 7.55(s, IH), 7.80(d, J=6.75Hz, IH), 8.03(d, J=8.80Hz, 2H)
Example 16
Preparation of 4-chloromethyl-benzoic acid-7-acetylamino-l,2,3- trimethoxy-10-methylsulfonyl-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl- methyl-ester
0.12g of the title compound 16 (yields: 40%, as a yellow solid) was prepared by the same method of Example 15 except that 4-hydroxymethylthiocolchicine was used instead of 4-hydroxymethylcolchicine.
Η NMR (500 MHz, CDC13) δ :1.83-1.85(m, IH), 2.02(s, 3H), 2.19-2.21(m,
2H), 2.43(s, 3H), 2.92-2.94(m, IH), 3.67(s, 3H), 3.98(s, 3H), 4.00(s, 3H), 4.61(s,
2H), 4.64-4.66(m, IH), 5.41(d, J=11.15Hz, IH), 5.46(d, J= 11.15Hz, IH), 7.06 (d, J= 10.56Hz, IH), 7.27(d, J= 9.39Hz, IH), 7.38(s, IH), 7.45(d, J=8.51Hz, 2H), 7.50 (d,
J=7.33Hz, IH), 8.03(d, J=8.51Hz, 2H)
Example 17
Preparation of 4-chlorobutyric acid-7-acetylamino-l,2,3,10-tetramethoxy -9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl-methyl ester
0.13g of the title compound 17 (yields: 92%, as a yellow solid) was prepared by the same method of Example 15 except that 4-chlolobutyryl chloride was used instead of 4-chloromethylbenzoyl chloride.
1H NMR (500 MHz, CDC13) δ :1.91-1.93(m, IH), 2.00(s, 3H), 2.10-2.12(m, 2H), 2.23-2.25(m, 2H), 2.55(t, J=7.04Hz, 2H), 2.82- 2.84(m, IH), 3.63(t, J=6.16Hz, 2H), 3.65(s, 3H), 3.96(s,3H), 3.98(s, 3H), 4.03(s, 3H), 4.63-4.66(m, IH), 5.12(d, J=12.91Hz, IH), 5.27(d, J=12.91Hz, IH), 6.90(d, J=11.15Hz, IH), 7.30(d, J=9.68Hz, IH), 7.61(s,lH), 8.21(d, J=6.45Hz, IH)
Example 18
Preparation of 4-chloro-butyric acid-7-acetylamino-l,2,3-trimethoxy-10- methyl-sulfonyl-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl-methyl ester
0.13g of the title compound 18 (yields: 92%, as a yellow solid) was prepared by the same method of Example 15 except that 4-hydroxymethyl thiocolchicine and 4-chlorobutyryl chloride were used instead of 4-hydroxymethyl colchicine and 4-chloromethylbenzoyl chloride, respectively.
1H NMR (500 MHz, CDC13) δ :1.85-1.87(m, IH), 2.03(s, 3H), 2.12-2.14(m, 2H), 2.19-2.21(m, 2H), 2.45(s, 3H), 2.53(t, J=7.33Hz, 2H), 2.81-2.83(m, IH), 3.60(t, J=6.75Hz, 2H), 3.65(s, 3H), 3.96 (s,3H), 3.98(s, 3H), 4.64-4.66(m, IH), 5.17(d, J=12.03Hz, IH), 5.26(d, J=12.03Hz, IH), 7.07(d, J=10.56Hz, IH), 7.26(d, J=10.27Hz, IH), 7.37(s,lH), 7.40(d, J=7.04Hz, IH) Example 19 Preparation of 4-nitrooxymethyl-benzoic acid-7-acetylamino-l, 2,3,10-tetra- methoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-4-yl-methyl-ester
0.084g (0.14mmol) of the compound 15 obtained from the Example 15 (0.14mmol) was placed into the flask and then dissolved by adding 5 ml of acetone. After adding 0.086g of sodium iodide(0.057mmol) dropwisely thereto, the mixture was stirred at 40-50 °C for 12 hours, extracted with chloroform, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. Concentrated resulting compound and 2 ml acetonitrile were placed with a 25ml flask and 0.09g (0.53mmol) of silver nitrate was added thereto. The mixture was stirred for 12 hours, and the solvent was filtered and removed under reduced pressure. The resulting compound was purified by column chromatography (ethylacetate: methanol = 85:15) to obtain 40mg of the title compound 19 (yields of second step reaction: 46%) as a yellow solid.
Η NMR of compound 19: Η NMR (500 MHz, CDC13) δ :1.86-1.88(m, IH), 1.77(s, IH), 2.20-2.22(m,
2H), 2.92-2.94(m, IH), 3.67(s, 3H), 3.98(s, 3H), 3.99(s,3H), 4.00(s,3H), 4.67-4.69(m, IH), 5.39-5.47(m, 2H), 5.47(s, 2H), 6.86(d, J=10.85Hz, IH), 7.30(d, J=10.85Hz, IH), 7.47(d, J=8.51Hz, 2H), 7.54(s, IH), 7.76(d, J=7.04Hz, IH), 8.01(d, J=8.51Hz, 2H) Example 20 Preparation of 4-nitrooxymethyl-butyric acid-7-acetylamino- 1,2,3
-trimethoxy-10-methylsulfonyl-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-4-yl- methyl-ester
70mg of the title compound 20 (yields of second step reaction: 39%, as a yellow solid) was prepared by the same method of the Example 19 through
4-iodomethyl-butyric acid-7-acetylamido- 1 ,2,3-trimethoxy- 10-methylsulfonyl-9-oxo- 5,6,7,9-tetrahydro-benzo[a]heptarene-4-yl-methyl-ester (intermediate compound 8) except for using compound 18 obtained from the Example 18.
Η NMR of intermediate compound 8:
1H NMR (500 MHz, CDC13) δ :1.82-1.84(m, IH), 2.02(s, 3H), 2.13-2.15(m,
4H), 2.44(s, 3H), 2.47(t, J=7.04Hz, 2H), 2.82-2.84(m, IH), 3.24(t, J=6.75Hz, 2H), 3.65(s, 3H), 3.95(s,3H), 3.97(s, 3H), 4.63-4.65(m, IH), 5.17(d, J=11.73Hz, IH),
5.25(d, J=l 1.73Hz, IH), 6.65(d, J=7.33Hz, IH), 7.05(d, J=10.27Hz, IH), 7.23(d,
J=10.27Hz, IH), 7.27(s,lH)
Η NMR of compound 20:
Η NMR (500 MHz, CDC13) δ :1.90-1.92(m, IH), 2.02(s, 3H), 2.08- 2.10(m, 2H), 2.18-2.20(m, 2H), 2.45(s, 3H), 2.49(t, J=7.04Hz, 2H), 2.81-2.83(m, IH), 3.65(s,
3H), 3.96(s,3H), 3.97(s, 3H), 4.54(t, J=6.45Hz, 2H), 4.65-4.67(m, IH), 5.18(d,
J=11.73, IH), 5.27(d, J=11.73Hz, 2H), 7.10(d, J=11.15Hz, IH), 7.26(d, J=9.68Hz,
IH), 7.45(s,lH), 8.00(d, J=7.04Hz, IH)
Example 21 Preparation of 4-nitrooxymethyl-benzoic acid-7-acetylamino- 1,2,3
-trimethoxy-10-methylsulfonyl-9-oxo-5,6,7,9,-tetrahydro-benzo[a]heptarene-4-yl- methyl- ester
70mg of the title compound 21 (yields of second step reaction: 45%, as a yellow solid) was prepared by the same method of the Example 19 through
4-iodomethyl-benzoic acid-7-acetylamino- 1 ,2,3-trimethoxy- 10-methylsulfonyl-9- oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-4-yl-methyl-ester(intermediate compound
9) except for using compound 16 obtained from the Example 16.
Η NMR of intermediate compound 9:
1H NMR (500 MHz, CDC13) δ :1.74-1.76(m, IH), 2.23(s, 3H), 2.13- 2.15(m,
IH), 2.23-2.25(m, IH), 2.43(s, 3H), 2.92-2.94(m, IH), 3.66 (s, 3H), 3.97(s,3H), 3.99(s, 3H), 4.45(s, 2H), 4.66-4.68(m.lH), 5.40(d, J=12.03Hz, IH), 5.45(d,
J=12.03Hz, IH), 6.35(d, J=6.45Hz, IH), 7.05(d, J=9.97Hz, IH), 7.24(d, J=7.24Hz,
IH), 7.22(s, IH), 7.41(d, J=8.51Hz, 2H), 7.96(d, J=8.51Hz, 2H)
Η NMR of compound 21:
Η NMR (500 MHz, CDC13) δ :1.79-1.81(m, IH), 2.00(s, 3H), 2.13- 2.15(m, IH), 2.23-2.25(m, IH), 2.43(s, 3H), 2.92-2.94(m, IH), 3.67 (s,3H), 3.97(s,3H), 3.99(s, 3H), 4.66-4.68(m.lH), 5.42-5.47(m, 2H), 5.47(s, 2H), 6.95(d, J=6.75Hz, IH), 7.05(d, J=10.56Hz, IH), 7.24(d, J=10.85Hz, IH), 7.30(s, IH), 7.46(d, J=8.21Hz, 2H), 8.07(d, J=8.21Hz, 2H)
Example 22 Deacetamidocolchicine-7-ol
2.29g (6.426mmol) of colchicine, 20ml of methanol and 20ml of chloromethane were placed into a 100ml flask and cooled to -78 °C . To the mixture, 0.729g (19.278mmol) of sodium borohydride was added and the mixture was warmed to 0- -20 °C with stirring for 5 hours. The solution was acidified with 50% acetic acid and extracted with chloroform, and then dried over sodium sulfate, filtered and the solvent was removed under reduced pressure. The concentrated resulting compound was recrystallized (methanol/ethylether) to obtain the title compound 22(2.0844g, 90.5%).
Η NMR (500 MHz, CDC13): δ 1.80-1.83 (m, IH), 2.40-2.49 (m, 3H), 3.26 (br, IH), 3.60 (s, 3H), 3.90 (s, 3H), 3.91 (s, 3H), 3.97 (s, 3H), 4.46-4.49 (m, IH), 6.55 (s, IH), 6.79 (d, J = 11.0 Hz, IH), 7.18 (d, J = 11.0 Hz, IH), 7.94 (s, IH)
Example 23
Preparation of (-)-4,7,7-trimethyl-3-oxo-2-oxa-bicyclo[2.2. l]heptane-l- carboxylic acid-l,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptalene -7-yl ester and (+)-4,7,7-trimethyl-3-oxo-2-oxa-bicyclo[2.2.1]heptane-l-carboxcylic acid-l,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptalene-7-yl ester
Distilled Pyridine
0.5098g (1.4225 mmol) of deacetamidocolchicine-7-ol was placed into a 10ml flask and 8ml of distilled pyridine was added thereto. To the mixture, (-)-camphanic chloride (0.4007g, 1.8492mmol) was added. After 3 hours, IN hydrochloric acid was added to stop the reaction. The mixture was extracted with ethyl acetate for three times, and then dried over anhydrous sodium sulfate and filtered, and the compound was concentrated under reduced pressure. The concentrated compound was purified by column chromatography (ethyl acetate) to obtain the racemates of the title compounds 23(1) and 23(2). The racemates were separated by MPLC (ethano isopropyl alcohol = 9:1) to obtain the compounds 23(1) (184.6mg, 24.1%) and 23(2) (191.5mg, 25.0%).
Η NMR of compound 23(1):
Η NMR (500 MHz, CDC13): δ 0.95 (s, 3H), 1.10 (s, 3H), 1.11 (s, 3H),
1.71-1.74 (m, IH), 1.90-1.93 (m, IH), 2.05-2.13 (m, 2H), 2.14-2.43 (m, 2H),
2.49-2.52 (m, IH), 2.54-2.58 (m, IH), 3.66 (s, 3H), 3.92 (m, 3H), 3.94 (s, 3H),
3.99 (s, 3H), 5.41 (dd, J = 7.0, 11.0 Hz, IH), 6.56 (s, IH), 6.80 (d, J = 11.0 Hz,
IH), 7.28 (d, J = 11.0 Hz, IH), 7.40 (s, IH)
Η NMR of compound 23(2):
Η NMR (500 MHz, CDC13): δ 1.02 (s, 3H), 1.07 (s, 3H), 1.12 (s, 3H), 1.71-1.74 (m, IH), 1.92-1.95 (m, IH), 2.07-2.11 (m, 2H), 2.35-2.43 (m, IH), 2.48-2.56 (m, 3H), 3.66 (s, 3H), 3.91 (m, IH), 3.94 (s, 3H), 3.99 (s, 3H), 5.46 (dd, J = 7.0, 11.0 Hz, IH), 6.56 (s, IH), 6.80 (d, J = 11.0 Hz, IH), 7.28 (d, J = 11.0 Hz, IH), 7.36 (s, lH)
Example 24
(-)-deacetamidocolchicine-7-ol
184.6mg (0.3427mmol) of the compound 23(1) obtained from the Example 23 was placed into a 10ml flask, and 2.5ml of methanol and 2.5ml of chloroform were added thereto. The solution was cooled to -78 °C, and 1.71ml (3.4275mmol) of 2N aqueous solution of sodium hydroxide was added into the solution. The mixture was warmed to the room temperature and stirred for 3 hours. It was extracted with chloroform for three times and washed with water, and then dried over anhydrous sodium sulfate. The solvent in the mixture was removed under reduced pressure. The resulting concentrated product was recrystallized (ethyl acetate/hexane) to obtain the title product 24 (128.8mg, 100%).
25
[α ] ~D : -102.55 (CDC13, c = 6.875 x KTg ml) 1H NMR (500 MHz, CDC13): δ 1.80-1.83 (m, IH), 2.40-2.49 (m, 3H),
3.26 (br, IH), 3.60 (s, 3H), 3.90 (s, 3H), 3.91 (s, 3H), 3.97 (s, 3H), 4.46-4.49
(m, IH), 6.55 (s, IH), 6.79 (d, J = 11.0 Hz, IH), 7.18 (d, J = 11.0 Hz, IH),
7.94 (s, IH) Example 25
(+)-deacetamidocolchicine-7-ol
191.5mg (0.3556mmol) of the compound 23(2) obtained from the Example 23 was added to a 10ml flask, and 2.5ml of methanol and 2.5ml of chloromethane were added thereto. The temperature was cooled down to -78 °C, and 1.78ml (3.5560mmol) of 2N aqueous solution of sodium hydroxide was added thereto. The solution was warmed to the room temperature and stined for 3 hours. The mixture was extracted with chloroform for three times and washed with water, and dried over anhydrous sodium sulfate and the solvent was removed by concentrating under reduced pressure. The resulting concentrated product was recrystallized (ethyl acetate/hexane) to obtain the title product 25(127.4mg, 100%).
[α ] ^ : +113.10 (CDC13, c = 7.250 x 10-3g/ml)
Η NMR (500 MHz, CDC13): δ 1.80-1.83 (m, IH), 2.40-2.49 (m, 3H), 3.26 (br, IH), 3.60 (s, 3H), 3.90 (s, 3H), 3.91 (s, 3H), 3.97 (s, 3H), 4.46-4.49 (m, IH), 6.55 (s, IH), 6.79 (d, J = 11.0 Hz, IH), 7.18 (d, J = 11.0 Hz, IH), 7.94 (s, IH)
Example 26
(-)-3-chloromethyl-benzoic acid-l,2,3,10-tetramethoxy-9-oxo-5,6,7,9- tetrahydro-bezo[a]heptarene-7-yl ester
61.2mg (0.171mmol) of (-)-deacetamidocolchicine-7-ol was placed into a 25ml flask, and 1 ml of chloromethane and 2ml of THF were added thereto. To the solution, 26.7 μi (0.188mmol) of 3-chloromethylbenzoyl chloride was added dropwise and then llAμ (0.512mmol) of triethylamine was added. 4.2mg (0.0342mmol) of DMAP was added to the mixture and it was stirred at room temperature for 3 hours. The mixture was extracted with chloroform and dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting concentrated product was purified with column chromatography (ethyl acetate: methanol = 20:1) to obtain the title product 26 (60.0mg, 68.8%).
Η NMR (500 MHz, CDC13): δ 2.15-2.22 (m, IH), 2.45-2.64 (m, 3H), 3.70 (s, 3H), 3.92 (s, 3H), 3.96 (s, 3H), 3.99 (s, 3H), 4.62 (s , IH), 4.63 (s, IH), 5.57 (dd, J = 6.5, 5.5 Hz, IH), 6.59 (s, IH), 6.83 (d, J = 11.0 Hz, IH), 7.33 (d, J = 11.0 Hz, IH), 7.46 (t, J = 7.5 Hz, IH), 7.53 (s, IH), 7.62 (d, J = 7.5 Hz, IH), 8.03 (d, J = 7.5 Hz, IH), 8.06 (s, IH)
Example 27
(+)-3-chloromethyl-benzoic acid-l,2,3,10-tetramethoxy-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptaren-7-yl ester
The title compound 27(58.7mg, 67.4%) was obtained in accordance with the same method of the Example 26 except that (+)-deacetamidocolchicine-7-ol was used instead of (-)-deacetamidocolchicine-7-ol.
Η NMR (500 MHz, CDC13): δ 2.15-2.22 (m, IH), 2.45-2.64 (m, 3H), 3.70 (s, 3H), 3.92 (s, 3H), 3.96 (s, 3H), 3.99 (s, 3H), 4.62 (s , IH), 4.63 (s, IH), 5.57 (dd, J = 6.5, 5.5 Hz, IH), 6.59 (s, IH), 6.83 (d, J = 10.8 Hz, IH), 7.33 (d, J = 10.8 Hz, IH), 7.46 (t, J = 7.5 Hz, IH), 7.53 (s, IH), 7.62 (d, J = 7.5 Hz, IH), 8.03 (d, J = 7.5 Hz, IH), 8.06 (s, IH)
Example 28
(-)-3-iodomethyl-benzoic acid- 1,2,3, 10-tetramethoxy-9-oxo-5, 6,7,9- tetrahydro-benzo[a]heptarene-7-yl ester
47.5mg of the title product 28(67.1%) was prepared by the same method of the first step of the Example 4 except for using of compound 26 obtained in the Example 26.
1H NMR (600 MHz, CDC13): δ 2.05-2.22 (m, IH), 2.46-2.62 (m, 3H),
3.71 (s, 3H), 3.93 (s, 3H), 3.97 (s, 3H), 3.99 (s, 3H), 4.48 (d , J = 9.9 Hz , IH),
4.50 (d , J = 9.9 Hz , IH), 5.57 (dd, J = 6.5, 4.3 Hz, IH), 6.59 (s, IH), 6.82 (d,
J = 10.8 Hz, IH), 7.33 (d, J = 10.8 Hz, IH), 7.41 (t, J = 7.8 Hz, IH), 7.51 (s,
IH), 7.61 (d, J = 7.8 Hz, IH), 7.96 (d, J = 7.8 Hz, IH), 8.05 (s, IH)
Example 29
(+)-3-iodomethyl-benzoic acid-l,2,3,10-tetramethoxy-9-oxo-5,6,7,9- tetrahydro- benzo[a]heptarene-7-yl ester
47.5mg of the title compound 29 (68.6%) was prepared by the first step of the Example 4 except for using compound 27 obtained the Example 27.
1H NMR (600 MHz, CDC13): δ 2.05-2.22 (m, IH), 2.46-2.62 (m, 3H),
3.71 (s, 3H), 3.93 (s, 3H), 3.97 (s, 3H), 3.99 (s, 3H), 4.48 (d , J = 9.9 Hz , IH),
4.50 (d , J = 9.9 Hz , IH), 5.57 (dd, J = 6.5, 4.3 Hz, IH), 6.59 (s, IH), 6.82 (d,
J = 10.8 Hz, IH), 7.33 (d, J = 10.8 Hz, IH), 7.41 (t, J = 7.8 Hz, IH), 7.51 (s,
IH), 7.61 (d, J = 7.8 Hz, IH), 7.96 (d, J = 7.8 Hz, IH), 8.05 (s, IH) Example 30
(-)-3-nitrooxymethyl-benzoic acid- 1 ,2,3,10-tetramethoxy-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptaren-7-yl ester
34.4mg of the title compound 30 (81.2%) was prepared by the second step of the Example 4 except for using compound 28 obtained the Example 28.
Η NMR (500 MHz, CDC13): δ 2.10-2.22 (m, IH), 2.46-2.62 (m, 3H), 3.70 (s, 3H), 3.92 (s, 3H), 3.96 (s, 3H), 3.99 (s, 3H), 5.48 (s , 2H), 5.57 ( dd, J = 6.5, 4.5 Hz, IH), 6.59 (s, IH), 6.82 (d, J = 10.8 Hz, IH), 7.32 (d, J = 10.8 Hz, IH), 7.50 (s, IH), 7.51 (t, J = 7.8 Hz, IH), 7.63 (d, J = 7.8 Hz, IH), 8.09 (s, IH), 8.10 (d, J = 7.8 Hz, IH)
Example 31
(+)-3-nitrooxymethyl-benzoic acid- 1,2,3, 10-tetramethoxy-9-oxo-5, 6,7,9- tetrahydro-benzo[a]heptarene-7-yl ester
32.5mg of the title compound 31 (76.7%) was prepared by the second step of the Example 4 except for using compound 29 obtained the Example 29.
Η NMR (500 MHz, CDC13): δ 2.10-2.22 (m, IH), 2.46-2.62 (m, 3H), 3.70 (s, 3H), 3.92 (s, 3H), 3.96 (s, 3H), 3.99 (s, 3H), 5.48 (s , 2H), 5.57 ( dd, J = 6.5, 4.5 Hz, IH), 6.59 (s, lH), 6.82 (d, J = 10.8 Hz, IH), 7.32 (d, J = 10.8 Hz, IH), 7.50 (s, IH), 7.51 (t, J = 7.8 Hz, IH), 7.63 (d, J = 7.8 Hz, IH), 8.09 (s, IH), 8.10 (d, J = 7.8 Hz, IH)
Table 1 shows the list of the compounds synthesized in accordance with the method of the present invention in addition to the compounds of the above Examples Table 1A
Table lB(continued)
Table lC(continued
Table lD(continued)
Table lE(continued)
Example A: Anticancer effect test
To ascertain the anticancer effects of colchicine derivatives of the formula (I) according to the present invention, the following tests were carried out by sulforhodamine-B (SRB) cytotoxicity assay. For comparison of anticancer effects, conventional colchicine and taxol as an anticancer drug were used as control groups. Human tumor cells, including MCF-7 (human breast adenomatous tumor),
MCF-7/DOX (adriamycin resistant cell strain), MRS-SA (human uterine sarcoma), MES-SA/DX5 (adriamycin resistant cell strain), A 549 (human non-small cell lung), SKOV-3 (human ovarian), SKMEL-2 (human melanoma), XF-498 (human CNS), HCT-15 (human colon) were incubated at 37°C in the presence of 5% CO2 using a DMEM culture medium. The respective cells were seeded into each well of 96-well plates at a concentration of 2xl03~5><103 cells/well. After culturing for 24 hours, colchicine dissolved in dimethylsulfoxide (DMSO), a compound 6, a compound 12 and taxol were diluted, and further cultured for 72 hours. Each cell line of the resultant plates was fixed with trichloroacetic acid (TCA), stained with 0.4% SRB solution and rinsed with 1% acetic acid. Thereafter, the dye was dissolved in lOmM Tris base to measure the optical density (OD) at 520 nm. The measurement results are listed in Tables 2 and 3. Table 2
Table 3
When ED50 values of the compounds 6 and 12 were investigated for each cell line, the colchicine derivative of the present invention exhibited a higher anticancer effect even at a low concentration of 0.02 to 773 nM than the conventional colchicine and taxol as shown in Tables 2 and 3.
Example B: Mixed lymphocyte reaction (MLR) tests MLR tests were carried out to determine the immunosuppressive effect of an immunosuppressive candidate material. When a responding cell (BALB/c mouse spleen cell) and a stimulating cell (DBA/2 mouse spleen cell) were cultured separately, the cells grew little. On the contrary, when the cells were simultaneously cultured, the cells were proliferated due to induction of an antigen-antibody reaction. The proliferated cells were treated with the immunosuppressive candidate material for measurement of the proliferation inhibitory extent.
Responding cells (BALB/c mouse spleen cells) and stimulating cells (DBA/2 mouse spleen cells) were respectively seeded into each well of 96-well plates at a concentration of 2 <10 cells/well for simultaneous culturing, and cyclosporin A (positive control group), colchicines, and colchicine derivatives of the present invention (compounds 6, 9, 10, 11 and 12) were treated. After culturing for 72 hours at a CO2 incubator, a 20 μi MTS solution was added to each well, followed by further culturing for 2 to 4 hours and measuring OD at 490 nm by using ELIS A. The results were shown in FIG. 1.
As shown in FIG. 1, it can be found that the colchicine derivatives according to the present invention, that is, the compounds 6, 9, 10, 11 and 12, suppressed growth of cells even at concentrations as low as 100 to 1000 nm and had a good immunosuppressive effect.
Example C: Immunosuppressive effect test using BALB/c mouse spleen cells
This test was carried out to determine the immunosuppressive effect of an immunosuppressive candidate material by checking anti-proliferation of T cells and B cells. To identify the proliferation inhibitory extent, responding cells (BALB/c mouse spleen cells) were treated with an immunosuppressive candidate material, lipopolysaccharide (LPS) as a B cell activator and concanvalin A (ConA) as a T cell activator. It is known that the responding cells treated with LPS induce proliferation of B cells and those treated with ConA induce proliferation of T cells.
Responding cells (BALB/c mouse spleen cells) were seeded into each well of 96-well plates at a concentration of 2*105 cells/well. Then, 20 μg/ml lipopolysaccharide, cyclosporin A (positive control group), and inventive colchicine derivatives (compounds 6, 10 and 12) were simultaneously treated, and 2 μg/ml of ConA, cyclosporin A (positive control sample), and inventive colchicine derivatives (compounds 6, 10 and 12) were simultaneously treated. After culturing for 72 hours at a CO2 incubator, a lOμA MTS solution was added to each well, followed by further culturing for 2 to 4 hours and measuring OD at 490 nm using ELISA. The measurement results are shown in FIGS. 2 and 3.
As shown in FIGS. 2 and 3, the colchicine derivatives according to the present invention suppressed proliferation of the B and T cells induced by LPS and ConA, respectively, in a concentration-dependent manner. The immunosuppressive effect of the colchicine derivatives according to the present invention was superior to that of cyclosporin A, a conventional immunosuppressive agent. In particular, the compounds 6 and 12 exhibited a remarkable anti-proliferous effect on B and T cells (FIG. 3). Example D: Toxicity assay
The colchicine derivatives according to the present invention were administered to ICR mice of 4-5 weeks old, weighing 18 to 20g, for acute (intravenous administration) toxicity assay and oral administration toxicity assay, as demonstrated in Tables 5 and 7. The same assays were carried out using colchicine as a control group (see Tables 4 and 6).
Example E: Reverse mutation assay by using bacteria
This assay is carried out to test mutagenicity of chemical substances using histidine auxotrophic strains that are one of Salmonella Typhimurium mutants. In the assay, artificially induced mutants (histidine auxotrophic mutants) are cultured in a histidine-free culture medium. When various mutagens are added to the culture medium, only revertants generated by reverse mutation survive. Thus generated colonies and spontaneously induced revertants are compared for detection of mutation.
Ames test was carried out using WP2 uvrA strains (tryptophan auxotrophic strains) of TA100, TA1535, TA98, TA1537 and Escherichia coli. Mutagens used as positive control samples were 0.5 g/plate of sodium azide, 0.5 g/plate of 4NQO (4-nitroquinolin-l -oxide), 50 g/plate of 9-AA (9-aminoacridine). Colchicine and colchicine derivative (Compound 6) according to the present invention were treated at amounts of 0, 317.5, 625, 1250, 2500 and 5000 g/plate, and incubated in the presence (+) and absence (-) of microsomal polysubsfrate oxygenases (S-9 mixture) at 37 °C for 48 hours. After incubation, the number of revertant colonies was counted. 3 plates were prepared for each test and the average was calculated. The results are demonstrated in Tables 8 and 9.
Table 8
Table 9
As shown in Tables 8 and 9, the colchicine derivatives according to the present invention induced noticeably fewer revertant colonies than the positive control groups, implying no significant hazard as mutagens.
Industrial Applicability
As described above, the novel colchicine derivative of the formula (I) according to the present invention or pharmaceutically acceptable salts thereof are superior to conventional colchicine in view of anticancer, anti-proliferous and immunosuppressive effects, and have less likelihood of toxicity and less hazard as mutagen than conventional colchicine.

Claims

What claimed is:
1. Colchicine derivative of the following formula (I) and pharmaceutically acceptable salts thereof:
wherein: when R, is N(R6)C(Xι)-A, X2C(Xι)-A, N(Rβ)-A, N(A)2 or X2-A, R2 is X3R7 or N(R7)2, R3 and R4 are independently hydrogen or a methyl group, R5 is hydrogen, a methyl group, or CH2X4R7, wherein R$ and R7 are independently hydrogen or a lower alkyl group, and X, Xj, X2, X3 and X4 are independently O or S; when R2 is N(R6)C(Xι)-A, X2C(Xι)-A, N(Re)-A, N(A)2 or X2-A, R, is
N(R6)COCH3, N(R6)COCF3, or NHC(O)OR8, R3 and R* are independently hydrogen or a methyl group, R5 is hydrogen, a methyl group, or CH2X4R7, wherein R6 and R are independently hydrogen or a lower alkyl group, R8 is a lower alkyl, alkenyl, or substituted or unsubstituted aryl, and X, Xi, X2 and X are independently O or S; when R3 and R4 are independently C(Xj)-A or -A, Rx is N(Re)COCH ,
N(R6)COCF3, or NHC(O)ORg, R2 is X3R7 or N(R7)2, R5 is hydrogen, a methyl group, or CH2XjR7, wherein Rβ and R7 are independently hydrogen or a lower alkyl group, R8 is a lower alkyl, alkenyl, or substituted or unsubstituted aryl, and X, Xi, X3 or t are independently O or S; when R5 is CH2X2C(X,)-A, R, is N(R6)COCH3, N(R6)COCF3 or NHC(O)OR8,
R2 is X3R7 or N(R7)2, R3 and R-j are independently hydrogen or a methyl group, wherein Rό and R are independently hydrogen or a lower alkyl group, R8 is a lower alkyl, alkenyl, or substituted or unsubstituted aryl group, and X, Xι,X2 and X3 are independently O or S, wherein A is represented by the formula (a), (b), (c), (d), (e), (f), (g), (h), (i) or 0):
— Y,-Hal (a) — Y,-ON02 (b)
-(CH-CHz-OJn,- Hal (C) -(CH-CH2-0)n,-N02 (d)
— (CH2)n2-CH-(CH2)n3-CH3 (e) -(CH2)n2-CH-(CH2)n3-CHj (0 Hal ONO,
wherein Y] is a Ci to Cι0 straight chain or branched alkyl, preferably, a C2 to C5 straight chain or branched alkyl or a substituted C5 to C7 cycloalkyl group; Hal is halogen, for example, F, Cl, Br or I; R is hydrogen or a lower alkyl group; n] is an integer from 1 to 6, preferably from 2 to 4; n2 and n3 are independently an integer from 1 to 5, preferably from 1 to 3; iu is an integer from 0 to 3; and n5 is an integer
2. A compound selected from the group consisting of: 4-chloro-N-(l,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl)- butylamide;
4-chloromethyl-N-(l ,2,3, 10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-benzamide;
3-chloromethyl-N-(l ,2,3, 10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-benzamide; 4-iodo-N-(l ,2,3, 10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl)- butylamide;
4-nitrooxy-N-( 1 ,2,3 , 10-tetramethoxy-9-oxo-5 ,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-butylamide; 4-iodomethyl-N-(l,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-benzamide;
4-nitrooxymethyl-N-(l,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-benzamide;
3-iodomethyl-N-( 1 ,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-benzamide;
3-nitrooxymethyl-N-(l,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-benzamide;
4-chloro-N-( 1 ,2,3 -trimethoxy- 10-methylsulfanyl-9-oxo-5 ,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-butylamide; 4-chloromethyl-N-(l,2,3-trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-7-yl)-benzamide;
3-chloromethyl-N-(l,2,3-trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-7-yl)-benzamide;
4-iodo-N-(l,2,3-trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-butylamide;
4-nitrooxy-N-(l,2,3-trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-butylamide;
4-iodomethyl-N-(l,2,3-trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-benzamide; 4-nitrooxymethyl-N-( 1,2,3 -trimethoxy- 10-methylsulfanyl-9-oxo-5, 6,7,9- tetrahydro-benzo[a]heptarene-7-yl)-benzamide;
3-iodomethyl-N-(l,2,3-trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-benzamide;
3-nitrooxymethyl-N-(l,2,3-trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-7-yl)-benzamide; N-(7-acetylamino- 1 ,2,3 -trimethoxy-9-oxo-5 ,6,7,9-tetrahydro-benzo [a]heptarene- 10- yl)-4-chloromethyl-benzamide;
N-(7-acetylamino-l,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-10- yl)-4-iodomethyl-benzamide; N-(7-acetylamino- 1,2,3 -trimethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene- 10- yl)-4-nitrooxymethyl-benzamide;
4-chloromethyl-benzoic acid 7-acetylamino- 1,2,3, lO-tetramethoxy-9-oxo-
5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl-methyl ester;
4-chloromethyl-benzoic acid 7-acetylamino- 1 ,2,3-trimethoxy- 10-methylsulfanyl- 9-oxo-5, 6,7,9-tetrahydro-benzo[a]heptarene-7-yl-methyl ester;
4-chloro-butyric acid 7-acetylamino- 1,2,3, 10-tetramethoxy-9-oxo-5, 6,7,9- tetrahydro-benzo[a]heptarene-7-yl-methyl ester;
4-chloro-butyric acid 7-acetylamino- 1 ,2,3-trimethoxy- 10-methylsulfanyl-9-oxo-
5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl-methyl ester; 4-nitrooxymethyl-benzoic acid 7-acetylamino-l,2,3,10-tetramethoxy-9-oxo-
5,6,7,9-tetrahydro-benzo[a]heptarene-4-yl-methyl ester;
4-iodomethyl-butyric acid 7-acetylamino- 1 ,2,3-trimethoxy- 10-methylsulfanyl-9- oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-4-yl-methyl ester;
4-nitrooxymethyl-butyric acid 7-acetylamino- 1,2,3-trimethoxy- 10- methylsulfanyl-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-4-yl-methyl ester;
4-iodomethyl-benzoic acid 7-acetylamino- 1 ,2,3-trimethoxy- 10-methylsulfanyl-9- oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-4-yl-methyl ester;
4-nitrooxymethyl-benzoic acid 7-acetylamino- 1,2,3 -trimethoxy- 10- methylsulfanyl-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-4-yl-methyl ester; (-)-3-chloromethyl-benzoic acid 1,2,3, 10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl ester;
(+)-3-chloromethyl-benzoic acid 1,2,3, 10-tetramethoxy-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-7-yl ester;
(-)-3-iodomethyl-benzoic acid 1,2,3, 10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl ester; (+)-3-iodomethyl-benzoic acid 1, 2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl ester;
(-)-3-nitrooxymethyl-benzoic acid 1,2,3, 10-tetramethoxy-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-7-yl ester; (+)-3-nitrooxymethyl-benzoic acid 1, 2,3,10-tetramethoxy-9-oxo-5, 6,7,9- tetrahydro-benzo[a]heptarene-7-yl ester;
4-chloro-butyric acid 7-acetylamino-l,2,10-trimethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-3-yl ester;
4-chloromethyl-benzoic acid 7-acetylamino-l,2,10-trimethoxy-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-3-yl ester;
3-chloromethyl-benzoic acid 7-acetylamino-l,2,10-trimethoxy-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-3-yl ester;
4-nitrooxy-butyric acid 7-acetylamino- 1,2,10-trimethoxy-9-oxo-5, 6,7,9- tetrahydro-benzo[a]heptarene-3-yl ester; 4-nitrooxymethyl-benzoic acid 7-acetylamino- 1,2,10-trimethoxy-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-3-yl ester;
3-nitrooxymethyl-benzoic acid 7-acetylamino- 1,2,10-trimethoxy-9-oxo-5, 6,7,9- tetrahydro-benzo[a]heptarene-3-yl ester;
4-chloro-N-methyl-N-(l,2,3-trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-7-yl)-butylamide;
4-nitrooxymethyl-N-methyl-N-(l,2,3-trimethoxy-10-methylsulfanyl-9-oxo-
5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl)-benzamide;
3-nitrooxymethyl-N-methyl-N-( 1 ,2,3-trimethoxy- 10-methylsulfanyl- 9-oxo-
5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl)-benzamide; (-)-3-chloromethyl-benzoic acid 1 ,2,3-trimethoxy- 10-methylsulfenyl-9-oxo-
5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl ester;
(+)-3-chloromethyl-benzoic acid 1,2,3 -trimethoxy- 10-methylsulfenyl-9-oxo-
5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl ester;
(-)-3-nitrooxymethyl-benzoic acid l,2,3-trimethoxy-10-methylsulfenyl-9-oxo- 5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl ester; (+)-3-nitrooxymethyl-benzoic acid 1 ,2,3-trimethoxy- 10-methylsulfenyl-9-oxo-
5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl ester;
4-chloro-N-methyl-N-( 1,2,3, 10-tetramethoxy-9-oxo-5, 6,7,9-tetrahydro- benzo[a]heptarene-7-yl) butylamide; 4-chloromethyl-N-methyl-N-(l,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl) benzamide;
N-methyl-4-nitrooxy-N-(l,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl) butylamide;
N-methyl-4-nitrooxymethyl-N-(l,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydro-benzo [a]heptarene-7-yl) benzamide;
4-nitrooxy-butyric acid 7-acetylamino- 1,2,3, 10-tetramethoxy-9-oxo-5, 6,7,9- tetrahydro-benzo[a]heptarene-4-yl methyl ester;
4-chloro-N-(l,2,3-trimethoxy-4-methoxymethyl-10-methylsulfanyl-9-oxo-
5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl)-butylamide; 4-chloromethyl-N-(l,2,3-trimethoxy-4-methoxymethyl-10-methylsulfanyl-9- oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl)-benzamide;
4-nitrooxy-N-(l,2,3-trimethoxy-4-methoxymethyl-10-methylsulfanyl-9-oxo-
5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl)-butylamide;
4-nitrooxymethyl-N-( 1 ,2,3-trimethoxy-4-methoxymethyl- 10-methylsulfanyl-9- oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-7-yl)-benzamide;
N-(7-acetylamino- 1 ,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene- 10- yl)-3-chloromethyl-benzamide;
N-(7-acetylamino-l,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-10- yl)-3-nitrooxymethyl-benzamide; N-(7-acetylamino-l,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptarene-10- yl-4-chloro-butylamide;
4-chloromethyl-N-(10-dimethylamino-l,2,3-trimethoxy-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-7-yl)-benzamide;
N-( 10-dimethylamino- 1 ,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-4-nitrooxymethyl-benzamide; 3-chloromethyl-N-(10-dimethylamino-l,2,3-trimethoxy-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptarene-7-yl)-benzamide; N-( 10-dimethylamino- 1 ,2,3 -trimethoxy-9-oxo-5 ,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-3-nitrooxymethyl-benzamide; 4-chloro-N-(10-dimethylamino-l,2,3-trimethoxy-9-oxo-5,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-butyrilamide; and N-( 10-dimethylamino- 1 ,2,3 -trimethoxy-9-oxo-5 ,6,7,9-tetrahydro- benzo[a]heptarene-7-yl)-4-nitrooxy-butyrilamide, or pharmaceutically acceptable salts thereof.
3. A pharmaceutical composition comprising the colchicine derivative or its pharmaceutically acceptable salt according to claim 1 or 2.
4. An anticancer agent, an anti-proliferous agent, and an immunosuppressive agent comprising the colchicine derivative or its pharmaceutically acceptable salt according to claim 1 or 2 as an active ingredient.
5. A method for preparing a colchicine derivative of the formula (la) among the colchicine derivatives according to claim 1 comprising: reacting a compound of the formula (U) with a compound of the following formula (IU) or a compound of the following formula (V) for amidation to produce a compound of the following formula (IV) or a compound of the following formula (VI); and subjecting the compound of the formula (IV) or (VI) for nitration to convert the same into the colchicine derivative of the formula (la):
wherein, B is represented by the following formulas Cl and C2:
(wherein Ri through R5 and X are defined as in claim 1),
, and wherein Re is hydrogen or a lower alkyl group; Xi is O or S; Hal, Hah, and Hal2 are independently the same or different halogen; and Y is represented by the general formulas (a'), (b1), (c*), (d') or (e1):
-(CH-CH2-0)n,— ( 1)
R«
— (CH2)n2— CH-(CH2)n3-CH3 (c1)
wherein Y\ represents a Ci to do straight chain or branched alkyl, preferably, a C2 to C5 straight chain or branched alkyl, or a substituted C5 to C7 cycloalkyl, R8 represents hydrogen or a lower alkyl, ni is an integer from 1 to 6, preferably from 2 to
4, n2 and n3 are independently an integer from 1 to 5, preferably from 1 to 3, r is an integer from 0 to 3, and n5 is an integer from 1 to 6.
6. The method according to claim 5 comprising nitration of a compound represented by the formula (VII) prepared by converting hydrogen of alcohol in the compound of the formula (VI) into a leaving group before the nitration of the compound of the formula (VI) :
wherein B, R^, Xi and Y are defined as in claim 5, and L is a leaving group selected from a methanesulfonyl, 7-toluenesulfonyl or triflate.
7. A method for preparing a colchicine derivative of the formula (lb) among the colchicine derivatives according to claim 1 comprising: reacting the compound of the formula (El) with a compound of the formula
(VIH) or a compound of the formula (X) for amidation to produce a compound (IX) or a compound of the formula (XI); and subjecting the compound of the formula (IX) or the compound of the formula (XI) to nitration for converting the same into the compound of the formula (lb):
wherein B is defined as in claim 5,
Hat— Y-OH B'^Y'0"
X XI
, and wherein R , Y, and Hal are defined as in claim 5; Hah and Hal are independently the same or different halogen.
8. The method according to claim 7 comprising nitration of a compound represented by the formula (XIT) prepared by converting hydrogen of alcohol in the compound of the formula (XI) into a leaving group before nitration of the compound of the formula (XI):
wherein B, R , and Y are defined as in claim 7, and L is a leaving group selected from a methanesulfonyl, p-tohiene sulfonyl or triflate.
9. A method of preparing a colchicine derivative of the formula (Ic) among the colchicine derivatives according to claim 1 comprising: reacting a compound of the formula (XIH) with a compound of the formula
(UT) or a compound of the formula (XV) for esterification to produce a compound of the formula (XIV) or a compound of the formula (XVI); and subjecting the compound of the formula (XIV) or the compound of the formula (XVI) to nitration to produce the compound of the formula (Ic):
Ic wherein C is represented by the formula Cl, C2, C3, C4 or C5:
wherein Rx through R5 and X are defined as in claim 1,
XV XVI
, and wherein Xi and X2 are each independently O or S; Hal is halogen; Hah and Hal2 are independently the same or different halogen; and Y is represented by the general formulas (a1), (b')} (c'), (d') or (e1): — Yi- (a')
-(CH-CH O ,— (b1)
-(CH2)n2-CH-(CH2)n3-CH3 (c1)
wherein Yi represents a Ci to Cio straight chain or branched alkyl, preferably, a C2 to C5 straight chain or branched alkyl, or a substituted C5 to C7 cycloalkyl, R8 represents hydrogen or a lower alkyl, ni is an integer from 1 to 6, preferably from 2 to 4, n2 and n are independently an integer from 1 to 5, preferably from 2 to 4, at is an integer 0 to 3, and n5 is an integer from 1 to 6.
10. The method according to claim 9 comprising reacting the compound of the formula (XIH) with a compound of the formula (XVHI) having a hydroxy-protecting group to produce a compound of the formula (XIX) before esterification of the compound of the formula (XHT); and converting the compound of the formula (XIX) into a compound of the formula (XVI) by a deprotection:
wherein C, Xi and Y are defined as in claim 9; P represents a general hydroxy-protecting group selected among methoxymethyl or t-butyldimethylsilyl; and Rio is hydrogen or a lower alcohol of Ci to C3.
11. The method according to claim 9 or 10 comprising nitration of a compound of the formula (XVIT) prepared by converting hydrogen of alcohol in the compound of the formula (XVI) into a leaving group before nitration of the compound of the formula (XVI):
xvπ wherein C, Xj, X2 and Y are defined as in claim 9, and L represents a leaving group selected from methanesulfonyl, -toluene sulfonyl or triflate.
12. A method for preparing a colchicine derivative of the formula (Id) among the colchicine derivatives according to claim 1 comprising: reacting a compound of the formula (XX) with a compound of the formula
(VIH) or a compound of the formula (X) for amidation to produce a compound of the formula (XXI) or a compound of the formula (XXII); and subjecting the compound of the formula (XXI) or the compound of the formula (XXH) to nitration to convert the same into the colchicine derivative of the formula (Id):
wherein B is defined as in claim 5,
B- H
Hal— Y-OH Hal,-Y-Hal2 VTO XX
wherein Y and Hal is defined as in claim 5; Hah and Hal2 are independently the same or different halogen.
13. The method according to claim 12 comprising nitration of a compound of the formula (XXIH) prepared by converting each hydrogen of alcohols in the compound of the formula (XXH) into a leaving group before nitration of the compound of the formula (XXH):
wherein B and Y are defined as in claim 12, and L represents a leaving group selected from methanesulfonyl, -toluene sulfonyl or triflate.
14. A method of preparing a colchicine derivative of the formula (le) among the colchicine derivatives according to claim 1 comprising: reacting a compound of the formula (XHI) with a compound of the formula
(VIH) or a compound of the formula (X) for esterification to produce a compound of the formula (XXIV) or a compound of the formula (XXV); and subjecting the compound of the formula (XXIV) or the compound of the formula (XXV) to nitration to convert the same into the colchicine derivative of the formula (le):
C-XrY-ONO«, le wherein C is defined as in claim 9,
C-XjH
C-Xi-Y-Kal] CXrY-OH
XHV | imd XXV
wherein X2, Y, Hal, Hah and Hal2 are defined as in claim 9.
15. The method according to claim 14 comprising nitration of a compound of the formula (XXVI) prepared by converting hydrogen of alcohol in the compound of the formula (XXV) into a leaving group before nitration of the compound of the formula (XXV):
C-X Y-OL XXVI wherein C, X2 and Y are defined as in claim 14, and L represents a leaving group selected from methanesulfonyl, /?-toluene sulfonyl or triflate.
EP02733537A 2001-05-28 2002-05-27 A novel alkaloid derivative and a pharmaceutical composition containing the same Expired - Lifetime EP1404652B1 (en)

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EP1646608B1 (en) * 2003-06-25 2011-03-02 JE IL Pharmaceutical Co., Ltd. Tricyclic derivatives or pharmaceutically acceptable salts thereof, their preparations and pharmaceutical compositions containing them
AR047732A1 (en) * 2003-12-02 2006-02-15 Nicox Sa NITROOXIDERIVATES OF ANTIHIPERTENSIVE DRUGS
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EP2435403A4 (en) * 2009-05-27 2015-04-01 Takeda Pharmaceuticals Usa Inc Thiocolchicine derivatives, method of making and methods of use thereof
KR101077609B1 (en) * 2009-07-08 2011-10-27 제일약품주식회사 Process for preparing tricyclic derivatives
JP5829520B2 (en) * 2009-08-20 2015-12-09 国立大学法人 千葉大学 Colchicine derivatives
KR101343443B1 (en) * 2010-02-18 2013-12-19 재단법인 아산사회복지재단 Colchicine derivatives or phamarceutically acceptable salts thereof, process for preparation thereof and pharmaceutical composition containing the same
WO2016139303A1 (en) * 2015-03-03 2016-09-09 Universität Zu Köln Pharmaceutical composition for the therapy of diseases caused by highly proliferating cells
KR20250172901A (en) 2018-06-29 2025-12-09 알버타 헬스 서비시즈 Methods and uses of colchicine derivatives
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KR100600158B1 (en) 2006-07-12
EP1404652B1 (en) 2007-08-29
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